keratinocyte growth factor Search Results


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  • 93
    Thermo Fisher keratinocyte growth factor
    Maturation of the dermal equivalent is required to support to the growth and stratification of the epidermis in full thickness skin. (A) Differential infiltration of keratinocytes into the dermal compartment. Representative low and high magnification H E images of full thickness models generated by culturing the dermal component for: (a) 14 days and (b) 28 days before addition of keratinocytes to the surface of the dermal model. In both cases, the models were raised to the air–liquid interface for a further 14 days to promote differentiation. Multiple low magnification images of the same cross‐section were taken and stitched together using image j software to allow the visualisation of the full length of the full thickness skin models. (A,a) There is an absence of a continuous thick epidermal layer when the dermis is cultured for 14 days due to the invasion of keratinocytes into the dermal compartment. (A,b) A continuous epidermis is formed across the entire model when the dermal compartment is matured for 28 days and no invasion is observed. Scale bars: 200 μm in full‐length cross‐section (left, low magnification) and 100 μm (right, high magnification). (B) Photographs showing the surface of full thickness skin models within the well insert. The dermal component was initially cultured for either (a) 14 days or (b) 28 days prior to the seeding of keratinocytes and culturing for a further 14 days at the air–liquid interface. A hole in the epidermis is apparent in the 14‐day dermal model (white arrow, left), whereas the epidermis is complete and uniform across the well insert in the more mature 28‐day dermal model (right). (C) <t>Keratinocyte</t> infiltration decreases with the maturation of the dermal equivalent. The graph shows that the percentage depth of penetration of keratinocytes into the dermal construct decreased as the dermis was allowed to mature for longer periods of time. The data were generated by capturing multiple images and assessing areas of keratinocyte infiltration using image j software to determine the depth of penetration (data represent mean ± SEM , n = 3).
    Keratinocyte Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore keratinocyte growth factor
    Schematic diagram of the ftSE cultivation. ( a ) Punches are cut out of the Matriderm™ matrix and ( b ) degassed in an exsiccator. ( c ) Then the matrix is seeded with human dermal fibroblasts. ( d ) On day nine, the ftSE is transferred into a cell culture insert system. ( e ) After four days of cultivation, the skin equivalent is sealed with a fibrin gel. ( f ) One day later, normal human <t>keratinocytes</t> are seeded onto the top of the equivalent. ( g ) On day 20, the equivalents are lifted to the air-liquid interface and cultivated further for ~10 days.
    Keratinocyte Growth Factor, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Amgen keratinocyte growth factor
    Fibrin as a vehicle for in situ delivery of cultured <t>keratinocytes</t> onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .
    Keratinocyte Growth Factor, supplied by Amgen, used in various techniques. Bioz Stars score: 91/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher human keratinocyte growth supplement kit hkgs kit
    Fibrin as a vehicle for in situ delivery of cultured <t>keratinocytes</t> onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .
    Human Keratinocyte Growth Supplement Kit Hkgs Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher human keratinocyte growth factor
    Gene expression levels in <t>keratinocyte</t> cells after exposure to neem extract. The mRNA expression levels were normalized to GAPDH expression, and the relative gene expression levels in the cells at 2, 4, 8, and 24 h after initiation of extract exposure were compared to the corresponding levels for unexposed cells, whose levels were defined as 1.0.
    Human Keratinocyte Growth Factor, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 313 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Biovitrum palifermin
    Gene expression levels in <t>keratinocyte</t> cells after exposure to neem extract. The mRNA expression levels were normalized to GAPDH expression, and the relative gene expression levels in the cells at 2, 4, 8, and 24 h after initiation of extract exposure were compared to the corresponding levels for unexposed cells, whose levels were defined as 1.0.
    Palifermin, supplied by Biovitrum, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Swedish Orphan Biovitrum palifermin
    Figure 3a. Overall survival by <t>palifermin</t> use Figure 3b. Event-free survival by palifermin use
    Palifermin, supplied by Swedish Orphan Biovitrum, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    PeproTech keratinocyte growth factor
    Figure 3a. Overall survival by <t>palifermin</t> use Figure 3b. Event-free survival by palifermin use
    Keratinocyte Growth Factor, supplied by PeproTech, used in various techniques. Bioz Stars score: 96/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Journal of Biological Chemistry keratinocyte growth factor
    Figure 3a. Overall survival by <t>palifermin</t> use Figure 3b. Event-free survival by palifermin use
    Keratinocyte Growth Factor, supplied by Journal of Biological Chemistry, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher bpe
    Figure 3a. Overall survival by <t>palifermin</t> use Figure 3b. Event-free survival by palifermin use
    Bpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher keratinocyte sfm
    Figure 3a. Overall survival by <t>palifermin</t> use Figure 3b. Event-free survival by palifermin use
    Keratinocyte Sfm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1799 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cohesion Biosciences keratinocyte growth factor
    The effect of adipose tissue extracellular fraction (AT-Ex) on intracellular reactive oxygen species (ROS) clearing in normal human fibroblasts (NHF), normal human melanocytes (NHM), and normal human <t>keratinocytes</t> (NHK). a Cells were pretreated or post-treated with 2% (v/v) AT-Ex or matched plasma for 24 h and exposed to H 2 O 2 . Twenty-four hours after H 2 O 2 treatment, cell viability was evaluated by MTT assay. Experiments were performed in triplicate; statistical significance versus untreated control (Ctrl) is reported as * p
    Keratinocyte Growth Factor, supplied by Cohesion Biosciences, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    RayBiotech keratinocyte growth factor
    AT-RvD3 accelerates re-epithelialization in cutaneous excisional surgical injury and promotes generation of the mitogen <t>keratinocyte</t> growth factor in injured lung. A and B: Keratinocyte growth factor (KGF) was measured by enzyme-linked immunosorbent assay
    Keratinocyte Growth Factor, supplied by RayBiotech, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    rdi research diagnostics keratinocyte growth factor
    AT-RvD3 accelerates re-epithelialization in cutaneous excisional surgical injury and promotes generation of the mitogen <t>keratinocyte</t> growth factor in injured lung. A and B: Keratinocyte growth factor (KGF) was measured by enzyme-linked immunosorbent assay
    Keratinocyte Growth Factor, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    R&D Systems recombinant human kgf fgf 7 protein
    AT-RvD3 accelerates re-epithelialization in cutaneous excisional surgical injury and promotes generation of the mitogen <t>keratinocyte</t> growth factor in injured lung. A and B: Keratinocyte growth factor (KGF) was measured by enzyme-linked immunosorbent assay
    Recombinant Human Kgf Fgf 7 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BioMimetic Therapeutics keratinocyte growth factor
    Fibrin as a vehicle for in situ delivery of cultured <t>keratinocytes</t> onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .
    Keratinocyte Growth Factor, supplied by BioMimetic Therapeutics, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Maturation of the dermal equivalent is required to support to the growth and stratification of the epidermis in full thickness skin. (A) Differential infiltration of keratinocytes into the dermal compartment. Representative low and high magnification H E images of full thickness models generated by culturing the dermal component for: (a) 14 days and (b) 28 days before addition of keratinocytes to the surface of the dermal model. In both cases, the models were raised to the air–liquid interface for a further 14 days to promote differentiation. Multiple low magnification images of the same cross‐section were taken and stitched together using image j software to allow the visualisation of the full length of the full thickness skin models. (A,a) There is an absence of a continuous thick epidermal layer when the dermis is cultured for 14 days due to the invasion of keratinocytes into the dermal compartment. (A,b) A continuous epidermis is formed across the entire model when the dermal compartment is matured for 28 days and no invasion is observed. Scale bars: 200 μm in full‐length cross‐section (left, low magnification) and 100 μm (right, high magnification). (B) Photographs showing the surface of full thickness skin models within the well insert. The dermal component was initially cultured for either (a) 14 days or (b) 28 days prior to the seeding of keratinocytes and culturing for a further 14 days at the air–liquid interface. A hole in the epidermis is apparent in the 14‐day dermal model (white arrow, left), whereas the epidermis is complete and uniform across the well insert in the more mature 28‐day dermal model (right). (C) Keratinocyte infiltration decreases with the maturation of the dermal equivalent. The graph shows that the percentage depth of penetration of keratinocytes into the dermal construct decreased as the dermis was allowed to mature for longer periods of time. The data were generated by capturing multiple images and assessing areas of keratinocyte infiltration using image j software to determine the depth of penetration (data represent mean ± SEM , n = 3).

    Journal: Journal of Anatomy

    Article Title: Bioengineering the microanatomy of human skin

    doi: 10.1111/joa.12942

    Figure Lengend Snippet: Maturation of the dermal equivalent is required to support to the growth and stratification of the epidermis in full thickness skin. (A) Differential infiltration of keratinocytes into the dermal compartment. Representative low and high magnification H E images of full thickness models generated by culturing the dermal component for: (a) 14 days and (b) 28 days before addition of keratinocytes to the surface of the dermal model. In both cases, the models were raised to the air–liquid interface for a further 14 days to promote differentiation. Multiple low magnification images of the same cross‐section were taken and stitched together using image j software to allow the visualisation of the full length of the full thickness skin models. (A,a) There is an absence of a continuous thick epidermal layer when the dermis is cultured for 14 days due to the invasion of keratinocytes into the dermal compartment. (A,b) A continuous epidermis is formed across the entire model when the dermal compartment is matured for 28 days and no invasion is observed. Scale bars: 200 μm in full‐length cross‐section (left, low magnification) and 100 μm (right, high magnification). (B) Photographs showing the surface of full thickness skin models within the well insert. The dermal component was initially cultured for either (a) 14 days or (b) 28 days prior to the seeding of keratinocytes and culturing for a further 14 days at the air–liquid interface. A hole in the epidermis is apparent in the 14‐day dermal model (white arrow, left), whereas the epidermis is complete and uniform across the well insert in the more mature 28‐day dermal model (right). (C) Keratinocyte infiltration decreases with the maturation of the dermal equivalent. The graph shows that the percentage depth of penetration of keratinocytes into the dermal construct decreased as the dermis was allowed to mature for longer periods of time. The data were generated by capturing multiple images and assessing areas of keratinocyte infiltration using image j software to determine the depth of penetration (data represent mean ± SEM , n = 3).

    Article Snippet: HEKn were trypsinised, centrifuged, and 5 × 105 cells were re‐suspended in Keratinocyte Growth Medium supplemented with 5 ng mL−1 keratinocyte growth factor (KGF; Thermo Fisher Scientific), 140 μm CaCl2 (Sigma‐Aldrich, Dorset, UK), and 50 μg mL−1 ascorbic acid (Sigma‐Aldrich).

    Techniques: Generated, Software, Cell Culture, Construct

    CELF1 overexpression in melanoma within a distinct landscape of RBPs. a Frequency (%) of melanoma patients with mutations and/or copy number alterations (deletions or amplifications) in all known mRBPs ( n = 692), classified according to their RNA binding domains (Pfam nomenclature). Data were retrieved from the melanoma TCGA database using cBioPortal. BRAF and NRAS are included as references for classical melanoma-associated oncogenes (see Supplementary Table 1 for additional information). b Copy-number variation (CNV), gain and loss of the indicated RBPs in different tumor types showed on a bubble chart: BRCA breast invasive carcinoma, COAD colon adenocarcinoma, HNSC head and neck squamous cell carcinoma, KICH kidney chromophobe, KIRP kidney renal papillary carcinoma, LIHC liver hepatocellular carcinoma, LUAD lung adenocarcinoma, LUSC lung squamous cell carcinoma, PRAD prostate adenocarcinoma, SCME skin cutaneous melanoma. Circle size represents percentage of biopsies with CNV alterations. c CELF1 mRNA expression levels in TCGA melanoma samples categorized by disease stage. d Overall survival of melanoma patients separated as a function of high vs. low CELF1 mRNA (these considered with respect to the median expression of all data in the TCGA melanoma data set). Indicated are p -values estimated with the Gehan–Breslow–Wilcoxon test; p -(Log-rank test)=0.048. e Immunoblots illustrating CELF1 expression in normal skin cells (fibroblasts, keratinocytes and three independent pools of melanocytes) and the indicated melanoma cell lines. CELF2 was indicated as a reference for homolog ELAV-family member. f Representative micrographs of benign nevi, primary melanomas (vertical growth phase), and metastatic melanomas, showing a heightened CELF1 expression (brown staining) during tumor progression. Nuclei were counterstained in blue with hematoxilin. g Distribution of benign and malignant specimens (% of analyzed cases) separated as a function of CELF1 scoring, and pooled for vertical growth phase melanoma, skin metastases and lymph node metastases

    Journal: Nature Communications

    Article Title: Systems analysis identifies melanoma-enriched pro-oncogenic networks controlled by the RNA binding protein CELF1

    doi: 10.1038/s41467-017-02353-y

    Figure Lengend Snippet: CELF1 overexpression in melanoma within a distinct landscape of RBPs. a Frequency (%) of melanoma patients with mutations and/or copy number alterations (deletions or amplifications) in all known mRBPs ( n = 692), classified according to their RNA binding domains (Pfam nomenclature). Data were retrieved from the melanoma TCGA database using cBioPortal. BRAF and NRAS are included as references for classical melanoma-associated oncogenes (see Supplementary Table 1 for additional information). b Copy-number variation (CNV), gain and loss of the indicated RBPs in different tumor types showed on a bubble chart: BRCA breast invasive carcinoma, COAD colon adenocarcinoma, HNSC head and neck squamous cell carcinoma, KICH kidney chromophobe, KIRP kidney renal papillary carcinoma, LIHC liver hepatocellular carcinoma, LUAD lung adenocarcinoma, LUSC lung squamous cell carcinoma, PRAD prostate adenocarcinoma, SCME skin cutaneous melanoma. Circle size represents percentage of biopsies with CNV alterations. c CELF1 mRNA expression levels in TCGA melanoma samples categorized by disease stage. d Overall survival of melanoma patients separated as a function of high vs. low CELF1 mRNA (these considered with respect to the median expression of all data in the TCGA melanoma data set). Indicated are p -values estimated with the Gehan–Breslow–Wilcoxon test; p -(Log-rank test)=0.048. e Immunoblots illustrating CELF1 expression in normal skin cells (fibroblasts, keratinocytes and three independent pools of melanocytes) and the indicated melanoma cell lines. CELF2 was indicated as a reference for homolog ELAV-family member. f Representative micrographs of benign nevi, primary melanomas (vertical growth phase), and metastatic melanomas, showing a heightened CELF1 expression (brown staining) during tumor progression. Nuclei were counterstained in blue with hematoxilin. g Distribution of benign and malignant specimens (% of analyzed cases) separated as a function of CELF1 scoring, and pooled for vertical growth phase melanoma, skin metastases and lymph node metastases

    Article Snippet: Melanocytes were cultured in Medium 254 (Invitrogen) supplemented with 1% melanocyte growth factors (HMGS, Invitrogen), 0.2 mM CaCl2 , and 100 μg/mL penicillin/streptomycin; keratinocytes were cultured in Epilife Medium (Invitrogen) supplemented with 1% keratinocyte growth factors (HKGS, Invitrogen), 0.2 mM CaCl2 and 100 μg/mL penicillin/streptomycin and fibroblasts were cultured in DMEM, supplemented with 10% FBS, and 100 μg/mL penicillin/streptomycin.

    Techniques: Over Expression, RNA Binding Assay, Expressing, Western Blot, Staining

    ( A ) Immunoblot analysis of markers of cellular differentiation (Involucrin; Keratin 10) and proliferation (proliferating cellular nuclear antigen, PCNA; retinoblastoma tumor suppressor protein, pRB) in keratinocytes infected with a control virus (LXSN) or an HPV-16 E7-expressing retrovirus (E7) at 0, 1, 6, 24, and 72 hr after induction of differentiation. ( B ) Analysis of p21 Cip1 , expression, p21 Cip1 association with cdk2, and histone H1 (HH1) kinase activity associated with cdk2 at 0, 24, and 72 hr after induction of differentiation. To allow for quantitative comparisons, the different panels of LXSN and E7-expressing keratinocytes shown in this figure are derived from identical exposures of the same gel.

    Journal: Genes & Development

    Article Title: The human papillomavirus E7 oncoprotein can uncouple cellular differentiation and proliferation in human keratinocytes by abrogating p21Cip1-mediated inhibition of cdk2

    doi:

    Figure Lengend Snippet: ( A ) Immunoblot analysis of markers of cellular differentiation (Involucrin; Keratin 10) and proliferation (proliferating cellular nuclear antigen, PCNA; retinoblastoma tumor suppressor protein, pRB) in keratinocytes infected with a control virus (LXSN) or an HPV-16 E7-expressing retrovirus (E7) at 0, 1, 6, 24, and 72 hr after induction of differentiation. ( B ) Analysis of p21 Cip1 , expression, p21 Cip1 association with cdk2, and histone H1 (HH1) kinase activity associated with cdk2 at 0, 24, and 72 hr after induction of differentiation. To allow for quantitative comparisons, the different panels of LXSN and E7-expressing keratinocytes shown in this figure are derived from identical exposures of the same gel.

    Article Snippet: Differentiation was induced by allowing for keratinocyte growth in high-calcium-containing medium (2.0 m m calcium) consisting of Dulbecco’s modified Eagle medium (DMEM; GIBCO BRL) supplemented with 10% fetal bovine serum as described previously ( ).

    Techniques: Cell Differentiation, Infection, Expressing, Activity Assay, Derivative Assay

    Coimmunoprecipitation of HPV-16 E7 and p21 Cip1 in E7-expressing keratinocytes. ( A ) Immunoprecipitations were performed with E7-specific ( left ) and p21 Cip1 -specific ( right ) antibodies, resolved by SDS-PAGE and followed by immunoblot analysis with an E7-specific monoclonal antibody. ( B ) Immunoprecipitation was performed with a p21 Cip1 -specific ( left ) or an E7-specific ( right ) antibody, resolved by SDS-PAGE and followed by immunoblot analysis with a p21 Cip1 -specific antibody.

    Journal: Genes & Development

    Article Title: The human papillomavirus E7 oncoprotein can uncouple cellular differentiation and proliferation in human keratinocytes by abrogating p21Cip1-mediated inhibition of cdk2

    doi:

    Figure Lengend Snippet: Coimmunoprecipitation of HPV-16 E7 and p21 Cip1 in E7-expressing keratinocytes. ( A ) Immunoprecipitations were performed with E7-specific ( left ) and p21 Cip1 -specific ( right ) antibodies, resolved by SDS-PAGE and followed by immunoblot analysis with an E7-specific monoclonal antibody. ( B ) Immunoprecipitation was performed with a p21 Cip1 -specific ( left ) or an E7-specific ( right ) antibody, resolved by SDS-PAGE and followed by immunoblot analysis with a p21 Cip1 -specific antibody.

    Article Snippet: Differentiation was induced by allowing for keratinocyte growth in high-calcium-containing medium (2.0 m m calcium) consisting of Dulbecco’s modified Eagle medium (DMEM; GIBCO BRL) supplemented with 10% fetal bovine serum as described previously ( ).

    Techniques: Expressing, SDS Page, Immunoprecipitation

    Photomicrographs of undifferentiated ( A ) and differentiated ( B ) human keratinocytes, and undifferentiated ( C ) and differentiated ( D ) HPV-16 E7-expressing keratinocytes.

    Journal: Genes & Development

    Article Title: The human papillomavirus E7 oncoprotein can uncouple cellular differentiation and proliferation in human keratinocytes by abrogating p21Cip1-mediated inhibition of cdk2

    doi:

    Figure Lengend Snippet: Photomicrographs of undifferentiated ( A ) and differentiated ( B ) human keratinocytes, and undifferentiated ( C ) and differentiated ( D ) HPV-16 E7-expressing keratinocytes.

    Article Snippet: Differentiation was induced by allowing for keratinocyte growth in high-calcium-containing medium (2.0 m m calcium) consisting of Dulbecco’s modified Eagle medium (DMEM; GIBCO BRL) supplemented with 10% fetal bovine serum as described previously ( ).

    Techniques: Expressing

    Schematic diagram of the ftSE cultivation. ( a ) Punches are cut out of the Matriderm™ matrix and ( b ) degassed in an exsiccator. ( c ) Then the matrix is seeded with human dermal fibroblasts. ( d ) On day nine, the ftSE is transferred into a cell culture insert system. ( e ) After four days of cultivation, the skin equivalent is sealed with a fibrin gel. ( f ) One day later, normal human keratinocytes are seeded onto the top of the equivalent. ( g ) On day 20, the equivalents are lifted to the air-liquid interface and cultivated further for ~10 days.

    Journal: Bioengineering

    Article Title: Bioengineering of a Full-Thickness Skin Equivalent in a 96-Well Insert Format for Substance Permeation Studies and Organ-On-A-Chip Applications

    doi: 10.3390/bioengineering5020043

    Figure Lengend Snippet: Schematic diagram of the ftSE cultivation. ( a ) Punches are cut out of the Matriderm™ matrix and ( b ) degassed in an exsiccator. ( c ) Then the matrix is seeded with human dermal fibroblasts. ( d ) On day nine, the ftSE is transferred into a cell culture insert system. ( e ) After four days of cultivation, the skin equivalent is sealed with a fibrin gel. ( f ) One day later, normal human keratinocytes are seeded onto the top of the equivalent. ( g ) On day 20, the equivalents are lifted to the air-liquid interface and cultivated further for ~10 days.

    Article Snippet: For the ALI cultivation, the E3 medium, composed of the E2 medium supplemented with 50 µg/mL AAP and 5 ng/mL keratinocyte growth factor (Sigma-Aldrich, St. Louis, MO, USA), was used (800 µL medium, in the bottom well only), and the medium was changed three times a week.

    Techniques: Cell Culture

    Fibrin as a vehicle for in situ delivery of cultured keratinocytes onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Fibrin as a vehicle for in situ delivery of cultured keratinocytes onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .

    Article Snippet: KGF, keratinocyte growth factor; PCNA, proliferating cell nuclear antigen.

    Techniques: In Situ, Cell Culture, Mouse Assay

    Epidermal thickness increased with increasing concentration of fibrinogen. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (e) Epidermal thickness was calculated as the ratio of the surface area from the basal layer (BL) to the granular cell layer (GL) over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Epidermal thickness increased with increasing concentration of fibrinogen. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (e) Epidermal thickness was calculated as the ratio of the surface area from the basal layer (BL) to the granular cell layer (GL) over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Article Snippet: KGF, keratinocyte growth factor; PCNA, proliferating cell nuclear antigen.

    Techniques: Concentration Assay, Generated, Staining

    Fibrin can be employed as a vehicle to deliver keratinocytes onto dermis. Tissue-engineered skin substitutes were constructed by seeding keratinocytes in fibrin hydrogels containing fibrinogen (4 mg/mL), fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. At the indicated times after cell seeding, tissues were harvested. (a) Paraffin-embedded tissue sections were stained with hematoxylin and eosin. Frozen sections from tissues on day 10 postseeding were immunostained for (b) PCNA (green); or (c) K10 (red), and counterstained with the nuclear dye Hoechst (blue). bar = 50 μm. (d) .

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Fibrin can be employed as a vehicle to deliver keratinocytes onto dermis. Tissue-engineered skin substitutes were constructed by seeding keratinocytes in fibrin hydrogels containing fibrinogen (4 mg/mL), fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. At the indicated times after cell seeding, tissues were harvested. (a) Paraffin-embedded tissue sections were stained with hematoxylin and eosin. Frozen sections from tissues on day 10 postseeding were immunostained for (b) PCNA (green); or (c) K10 (red), and counterstained with the nuclear dye Hoechst (blue). bar = 50 μm. (d) .

    Article Snippet: KGF, keratinocyte growth factor; PCNA, proliferating cell nuclear antigen.

    Techniques: Construct, Staining

    Epidermal cell number increased with increasing concentration of fibrin. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL) and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface frozen sections were stained with the nuclear dye Hoechst. The dashed line denotes basement membrane (bar = 50 μm; original magnification 20×). (e) Cell density was measured as the number of cell nuclei per basement membrane length. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Epidermal cell number increased with increasing concentration of fibrin. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL) and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface frozen sections were stained with the nuclear dye Hoechst. The dashed line denotes basement membrane (bar = 50 μm; original magnification 20×). (e) Cell density was measured as the number of cell nuclei per basement membrane length. The symbol (*) denotes statistical significance ( p

    Article Snippet: KGF, keratinocyte growth factor; PCNA, proliferating cell nuclear antigen.

    Techniques: Concentration Assay, Generated, Staining

    The thickness of the neoepidermis correlated with the extent of vascularization of the underlying dermal support. In vivo neoepidermis was generated by seeding cultured keratinocytes within fibrin hydrogels onto the prevascularized dermis at 1 week postgrafting. (a) Two weeks after keratinocyte delivery, tissues were excised and paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (b) Epidermal thickness was calculated as the ratio of the surface area from the basal to the granular cell layers over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: The thickness of the neoepidermis correlated with the extent of vascularization of the underlying dermal support. In vivo neoepidermis was generated by seeding cultured keratinocytes within fibrin hydrogels onto the prevascularized dermis at 1 week postgrafting. (a) Two weeks after keratinocyte delivery, tissues were excised and paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (b) Epidermal thickness was calculated as the ratio of the surface area from the basal to the granular cell layers over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Article Snippet: KGF, keratinocyte growth factor; PCNA, proliferating cell nuclear antigen.

    Techniques: In Vivo, Generated, Cell Culture, Staining

    Gene expression levels in keratinocyte cells after exposure to neem extract. The mRNA expression levels were normalized to GAPDH expression, and the relative gene expression levels in the cells at 2, 4, 8, and 24 h after initiation of extract exposure were compared to the corresponding levels for unexposed cells, whose levels were defined as 1.0.

    Journal: Data in Brief

    Article Title: Fibroblast and keratinocyte gene expression following exposure to extracts of neem plant (Azadirachta indica)

    doi: 10.1016/j.dib.2017.12.035

    Figure Lengend Snippet: Gene expression levels in keratinocyte cells after exposure to neem extract. The mRNA expression levels were normalized to GAPDH expression, and the relative gene expression levels in the cells at 2, 4, 8, and 24 h after initiation of extract exposure were compared to the corresponding levels for unexposed cells, whose levels were defined as 1.0.

    Article Snippet: The cells were cultured in Medium 154 (Invitrogen) supplemented with human keratinocyte growth factor (HKGS; Invitrogen), according to the manufacturer's instructions.

    Techniques: Expressing

    In vitro release of cytokines, chemokines, and growth factors from primary adult human keratinocytes stimulated by TNF α . The amount of chemokines (a), of cytokines, and of growth factors (b) was measured by Multiplex system technology in the supernatants of primary adult human keratinocytes (pKC) plated at the same density (8–10 × 10 4 cells/cm 2 ) and grown in low Ca 2+ -containing medium (white bars). The treatment with 10 ng/ml TNF α was carried out for 48 hours in a serum-free medium when cells were 60–70% confluent (black bars). The data are expressed as pg/10 6 cells.

    Journal: Mediators of Inflammation

    Article Title: HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes

    doi: 10.1155/2017/7435621

    Figure Lengend Snippet: In vitro release of cytokines, chemokines, and growth factors from primary adult human keratinocytes stimulated by TNF α . The amount of chemokines (a), of cytokines, and of growth factors (b) was measured by Multiplex system technology in the supernatants of primary adult human keratinocytes (pKC) plated at the same density (8–10 × 10 4 cells/cm 2 ) and grown in low Ca 2+ -containing medium (white bars). The treatment with 10 ng/ml TNF α was carried out for 48 hours in a serum-free medium when cells were 60–70% confluent (black bars). The data are expressed as pg/10 6 cells.

    Article Snippet: The obtained primary cells were then plated on 6-well tissue culture plates (Costar), precoated with coating matrix (type I collagen, Gibco BRL), cultured using a specific keratinocyte-serum-free media at low Ca2+ concentration ( < 0.07 mM), and supplemented with human keratinocyte growth factors (Gibco BRL).

    Techniques: In Vitro, Multiplex Assay

    Proliferation and differentiation of HaCaT cells in low (0.07 mM) and high (1.8 mM) Ca 2+ -containing medium. (a) Proliferation of HaCaT cells plated at the same density (1.0 × 10 4 cells/cm 2 ) assessed by the MTT assay at 2, 6, 9, and 14 days of incubation. Values represent mean ± SD of three independent experiments. (b) Western blot analysis of the expression of keratinocyte (KC) differentiation markers (K10, K14, and involucrin) in HaCaT cells grown in low (a) and high (c) Ca 2+ -containing medium for 6 (A6 and C6) or 14 (A14 and C14) days. The relative intensities of band signals quantified by digital scanning densitometry are reported in the histogram; β -actin was used to normalize the results to protein content. This blot is a representative of three independent experiments. (c) Immunofluorescence staining of HaCaT cells, grown in low (A) and high (C) Ca 2+ -containing medium for 6 (A6 and C6) and 14 (A14 and C14) days, and of primary human KC, grown in low Ca 2+ -containing medium, with anti-K10 antibodies (magnification: 60x).

    Journal: Mediators of Inflammation

    Article Title: HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes

    doi: 10.1155/2017/7435621

    Figure Lengend Snippet: Proliferation and differentiation of HaCaT cells in low (0.07 mM) and high (1.8 mM) Ca 2+ -containing medium. (a) Proliferation of HaCaT cells plated at the same density (1.0 × 10 4 cells/cm 2 ) assessed by the MTT assay at 2, 6, 9, and 14 days of incubation. Values represent mean ± SD of three independent experiments. (b) Western blot analysis of the expression of keratinocyte (KC) differentiation markers (K10, K14, and involucrin) in HaCaT cells grown in low (a) and high (c) Ca 2+ -containing medium for 6 (A6 and C6) or 14 (A14 and C14) days. The relative intensities of band signals quantified by digital scanning densitometry are reported in the histogram; β -actin was used to normalize the results to protein content. This blot is a representative of three independent experiments. (c) Immunofluorescence staining of HaCaT cells, grown in low (A) and high (C) Ca 2+ -containing medium for 6 (A6 and C6) and 14 (A14 and C14) days, and of primary human KC, grown in low Ca 2+ -containing medium, with anti-K10 antibodies (magnification: 60x).

    Article Snippet: The obtained primary cells were then plated on 6-well tissue culture plates (Costar), precoated with coating matrix (type I collagen, Gibco BRL), cultured using a specific keratinocyte-serum-free media at low Ca2+ concentration ( < 0.07 mM), and supplemented with human keratinocyte growth factors (Gibco BRL).

    Techniques: MTT Assay, Incubation, Western Blot, Expressing, Immunofluorescence, Staining

    Morphology of in vitro generated skin microtissue ( a – c ) and tumour microtissue ( d – f ). ( a ) Haematoxylin and eosin (H E) staining of the physiological architecture of a human full-thickness skin microtissue on day 16 of air–liquid culture under static conditions showing the columnar basal keratinocyte layer, a stratified stratum corneum and the layers in-between. ( b ) Immunofluorescent TUNEL (green) and Ki67 (red) staining indicating proliferative keratinocyte stem cells in the basal layer (red) and the apoptotic (cornifying) cells (green) on the upper skin layer. ( c ) Collagen IV staining of the basement membrane of the skin. ( d ) H E staining of homogeneously distributed identical H292 cells throughout the tumour spheroid. ( e ) Immunofluorescent TUNEL (green) and Ki67 (red) staining indicating healthy tumour microtissue status containing a significant fraction of growing tumour cells (red). ( f ) Vimentin (red) staining of the tumour microtissue showing fibroblastic properties. ( b , c , e , f ) Nuclei stained with DAPI (blue). Scale bar: 100 µm.

    Journal: Scientific Reports

    Article Title: Simultaneous evaluation of anti-EGFR-induced tumour and adverse skin effects in a microfluidic human 3D co-culture model

    doi: 10.1038/s41598-018-33462-3

    Figure Lengend Snippet: Morphology of in vitro generated skin microtissue ( a – c ) and tumour microtissue ( d – f ). ( a ) Haematoxylin and eosin (H E) staining of the physiological architecture of a human full-thickness skin microtissue on day 16 of air–liquid culture under static conditions showing the columnar basal keratinocyte layer, a stratified stratum corneum and the layers in-between. ( b ) Immunofluorescent TUNEL (green) and Ki67 (red) staining indicating proliferative keratinocyte stem cells in the basal layer (red) and the apoptotic (cornifying) cells (green) on the upper skin layer. ( c ) Collagen IV staining of the basement membrane of the skin. ( d ) H E staining of homogeneously distributed identical H292 cells throughout the tumour spheroid. ( e ) Immunofluorescent TUNEL (green) and Ki67 (red) staining indicating healthy tumour microtissue status containing a significant fraction of growing tumour cells (red). ( f ) Vimentin (red) staining of the tumour microtissue showing fibroblastic properties. ( b , c , e , f ) Nuclei stained with DAPI (blue). Scale bar: 100 µm.

    Article Snippet: Upon arrival, skin tissues were transferred into 1 ml of E3 medium composed of EpiLife™ medium (MEPI500CA, Thermo Fisher Scientific Inc., Waltham, USA) supplemented with human keratinocyte growth supplement, including 0.2 ng/ml human epidermal growth factor (S0015, Thermo Fisher Scientific Inc.), 1.4 mM CaCl2 , 50 µg/ml ascorbic acid (Sigma–Aldrich, St. Louis, MO, USA) and 10 ng/ml keratinocyte growth factor (Preprotech, Hamburg, Germany).

    Techniques: In Vitro, Generated, Staining, TUNEL Assay

    Effects of cetuximab treatment on skin microtissue at day 5 of co-culture. ( a ) H E staining of untreated control group indicating maintained columnar basal keratinocyte layer. ( b ) Immunofluorescent TUNEL (green) and Ki67 (red, arrows) staining of untreated control group indicating proliferative keratinocytes in the basal layer. ( c ) H E staining of antibody-exposed group showing irregular arrangement of keratinocytes in the stratum basale. ( d ) Immunofluorescent TUNEL (green) and Ki67 (red) staining of antibody-exposed group demonstrating complete lack of proliferative cells. ( a , c ) Dotted lines (orange) framing keratinocytes inside the stratum basale. ( b , d ) Nuclei stained with DAPI (blue). Scale bar: 100 µm. ( e ) Gene expression of tumour necrosis factor alpha (TNFα) and Keratin 1 (KRT1) in skin microtissue of untreated control group (white bars) and antibody exposure group (black bars), n = 3. Data shown as mean + SD.

    Journal: Scientific Reports

    Article Title: Simultaneous evaluation of anti-EGFR-induced tumour and adverse skin effects in a microfluidic human 3D co-culture model

    doi: 10.1038/s41598-018-33462-3

    Figure Lengend Snippet: Effects of cetuximab treatment on skin microtissue at day 5 of co-culture. ( a ) H E staining of untreated control group indicating maintained columnar basal keratinocyte layer. ( b ) Immunofluorescent TUNEL (green) and Ki67 (red, arrows) staining of untreated control group indicating proliferative keratinocytes in the basal layer. ( c ) H E staining of antibody-exposed group showing irregular arrangement of keratinocytes in the stratum basale. ( d ) Immunofluorescent TUNEL (green) and Ki67 (red) staining of antibody-exposed group demonstrating complete lack of proliferative cells. ( a , c ) Dotted lines (orange) framing keratinocytes inside the stratum basale. ( b , d ) Nuclei stained with DAPI (blue). Scale bar: 100 µm. ( e ) Gene expression of tumour necrosis factor alpha (TNFα) and Keratin 1 (KRT1) in skin microtissue of untreated control group (white bars) and antibody exposure group (black bars), n = 3. Data shown as mean + SD.

    Article Snippet: Upon arrival, skin tissues were transferred into 1 ml of E3 medium composed of EpiLife™ medium (MEPI500CA, Thermo Fisher Scientific Inc., Waltham, USA) supplemented with human keratinocyte growth supplement, including 0.2 ng/ml human epidermal growth factor (S0015, Thermo Fisher Scientific Inc.), 1.4 mM CaCl2 , 50 µg/ml ascorbic acid (Sigma–Aldrich, St. Louis, MO, USA) and 10 ng/ml keratinocyte growth factor (Preprotech, Hamburg, Germany).

    Techniques: Co-Culture Assay, Staining, TUNEL Assay, Expressing

    Figure 3a. Overall survival by palifermin use Figure 3b. Event-free survival by palifermin use

    Journal: Bone marrow transplantation

    Article Title: Palifermin is efficacious in recipients of TBI-based but not chemotherapy-based allogeneic hematopoietic stem cell transplants

    doi: 10.1038/bmt.2012.115

    Figure Lengend Snippet: Figure 3a. Overall survival by palifermin use Figure 3b. Event-free survival by palifermin use

    Article Snippet: Palifermin (Kepivance, Swedish Orphan Biovitrium) is a recombinant human KGF that is more stable than endogenous KGF due to the removal of 23 amino acids from its N-terminus (product information).

    Techniques:

    Figure 2a. Cumulative incidence rate of aGVHD for all adults by palifermin use Figure 2b. Cumulative incidence rate of cGVHD for all adults by palifermin use

    Journal: Bone marrow transplantation

    Article Title: Palifermin is efficacious in recipients of TBI-based but not chemotherapy-based allogeneic hematopoietic stem cell transplants

    doi: 10.1038/bmt.2012.115

    Figure Lengend Snippet: Figure 2a. Cumulative incidence rate of aGVHD for all adults by palifermin use Figure 2b. Cumulative incidence rate of cGVHD for all adults by palifermin use

    Article Snippet: Palifermin (Kepivance, Swedish Orphan Biovitrium) is a recombinant human KGF that is more stable than endogenous KGF due to the removal of 23 amino acids from its N-terminus (product information).

    Techniques:

    The effect of adipose tissue extracellular fraction (AT-Ex) on intracellular reactive oxygen species (ROS) clearing in normal human fibroblasts (NHF), normal human melanocytes (NHM), and normal human keratinocytes (NHK). a Cells were pretreated or post-treated with 2% (v/v) AT-Ex or matched plasma for 24 h and exposed to H 2 O 2 . Twenty-four hours after H 2 O 2 treatment, cell viability was evaluated by MTT assay. Experiments were performed in triplicate; statistical significance versus untreated control (Ctrl) is reported as * p

    Journal: Stem Cell Research & Therapy

    Article Title: Adipose tissue-derived extracellular fraction characterization: biological and clinical considerations in regenerative medicine

    doi: 10.1186/s13287-018-0956-4

    Figure Lengend Snippet: The effect of adipose tissue extracellular fraction (AT-Ex) on intracellular reactive oxygen species (ROS) clearing in normal human fibroblasts (NHF), normal human melanocytes (NHM), and normal human keratinocytes (NHK). a Cells were pretreated or post-treated with 2% (v/v) AT-Ex or matched plasma for 24 h and exposed to H 2 O 2 . Twenty-four hours after H 2 O 2 treatment, cell viability was evaluated by MTT assay. Experiments were performed in triplicate; statistical significance versus untreated control (Ctrl) is reported as * p

    Article Snippet: Keratinocyte growth factor

    Techniques: MTT Assay

    AT-Ex increases the proliferation rate of normal cells. Normal human fibroblasts (NHF) ( a ), normal human keratinocytes (NHK) ( b ), normal human melanocytes (NHM) ( c ) and adipose-derived stem cell (ADSC) ( d ) cell cultures were grown in the presence of increasing doses (1%, 2%, 5%, and 10% v/v) of adipose tissue extracellular fraction (AT-Ex) or matched patient plasma for 3 and 6 days. Experiments were performed in full medium and starved medium. Cell proliferation was measured by MTT assay. Data represent the mean ± SD of 14 (NHK), 12 (NHM), 24 (NHF) and 5 (ADSCs) different experiments performed in duplicate; statistical significance versus untreated control is reported as * p

    Journal: Stem Cell Research & Therapy

    Article Title: Adipose tissue-derived extracellular fraction characterization: biological and clinical considerations in regenerative medicine

    doi: 10.1186/s13287-018-0956-4

    Figure Lengend Snippet: AT-Ex increases the proliferation rate of normal cells. Normal human fibroblasts (NHF) ( a ), normal human keratinocytes (NHK) ( b ), normal human melanocytes (NHM) ( c ) and adipose-derived stem cell (ADSC) ( d ) cell cultures were grown in the presence of increasing doses (1%, 2%, 5%, and 10% v/v) of adipose tissue extracellular fraction (AT-Ex) or matched patient plasma for 3 and 6 days. Experiments were performed in full medium and starved medium. Cell proliferation was measured by MTT assay. Data represent the mean ± SD of 14 (NHK), 12 (NHM), 24 (NHF) and 5 (ADSCs) different experiments performed in duplicate; statistical significance versus untreated control is reported as * p

    Article Snippet: Keratinocyte growth factor

    Techniques: Derivative Assay, MTT Assay

    The effect of adipose tissue extracellular fraction (AT-Ex) on cell migration. Scratch assay with Normal human fibroblasts (NHF) ( a ) and normal human keratinocytes (NHK) ( b ) were performed to monitor cell motility in vitro. Both cell types were grown to confluence and starved for 24 h before the monolayer were scratched and then exposed to AT-Ex (2%) or not (control cells; Ctrl). The rate of migration was evaluated after 18 h. The extent of the wound closure is presented as the percentage by which the original scratch width has decreased. Data presented are representative of four independent experiments. c Following 24 h treatment, AT-Ex induces downregulation of connexin-43 (Cx-43) gap junctional protein expression in NHF and increased expression of CD44, a membrane protein associated with augmented cell motility. d Western blot analysis shows increments in vascular endothelial growth factor (VEGF), N-cadherin, and fibronectin proteins. Images are representative of five independent experiments. Nuclei were labeled with bisbenzidine (DAPI). Original magnification 40×. Images are representative of several independent experiments

    Journal: Stem Cell Research & Therapy

    Article Title: Adipose tissue-derived extracellular fraction characterization: biological and clinical considerations in regenerative medicine

    doi: 10.1186/s13287-018-0956-4

    Figure Lengend Snippet: The effect of adipose tissue extracellular fraction (AT-Ex) on cell migration. Scratch assay with Normal human fibroblasts (NHF) ( a ) and normal human keratinocytes (NHK) ( b ) were performed to monitor cell motility in vitro. Both cell types were grown to confluence and starved for 24 h before the monolayer were scratched and then exposed to AT-Ex (2%) or not (control cells; Ctrl). The rate of migration was evaluated after 18 h. The extent of the wound closure is presented as the percentage by which the original scratch width has decreased. Data presented are representative of four independent experiments. c Following 24 h treatment, AT-Ex induces downregulation of connexin-43 (Cx-43) gap junctional protein expression in NHF and increased expression of CD44, a membrane protein associated with augmented cell motility. d Western blot analysis shows increments in vascular endothelial growth factor (VEGF), N-cadherin, and fibronectin proteins. Images are representative of five independent experiments. Nuclei were labeled with bisbenzidine (DAPI). Original magnification 40×. Images are representative of several independent experiments

    Article Snippet: Keratinocyte growth factor

    Techniques: Migration, Wound Healing Assay, In Vitro, Expressing, Western Blot, Labeling

    AT-RvD3 accelerates re-epithelialization in cutaneous excisional surgical injury and promotes generation of the mitogen keratinocyte growth factor in injured lung. A and B: Keratinocyte growth factor (KGF) was measured by enzyme-linked immunosorbent assay

    Journal: The American Journal of Pathology

    Article Title: Resolvin D3 and Aspirin-Triggered Resolvin D3 Are Protective for Injured Epithelia

    doi: 10.1016/j.ajpath.2016.03.011

    Figure Lengend Snippet: AT-RvD3 accelerates re-epithelialization in cutaneous excisional surgical injury and promotes generation of the mitogen keratinocyte growth factor in injured lung. A and B: Keratinocyte growth factor (KGF) was measured by enzyme-linked immunosorbent assay

    Article Snippet: Keratinocyte growth factor (KGF; alias FGF7) was measured in BALF by enzyme-linked immunosorbent assay, per manufacturer's instructions (RayBiotech, Norcross, GA).

    Techniques: Enzyme-linked Immunosorbent Assay

    Fibrin as a vehicle for in situ delivery of cultured keratinocytes onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Fibrin as a vehicle for in situ delivery of cultured keratinocytes onto the prevascularized dermal substrate. Athymic mice ( n = 6 per group) were subjected to full thickness, excisional wound (including the panniculous carnosus) of ∼2.5 cm 2 on their dorsum, and the skin was replaced with acellular human dermis of equal size. Acellular human dermis remained untreated (control) or was infiltrated with fibrin gel alone (FG) or in combination with conditioned medium (FG + CM). One week postgrafting cultured keratinocytes were delivered directly onto the dermis in a fibrin hydrogel and covered with a moist Telfa dressing for 3 days before they were exposed to air (a) . Neoepidermis at 2 weeks postseeding (b) .

    Article Snippet: Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo.

    Techniques: In Situ, Cell Culture, Mouse Assay

    Epidermal thickness increased with increasing concentration of fibrinogen. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (e) Epidermal thickness was calculated as the ratio of the surface area from the basal layer (BL) to the granular cell layer (GL) over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Epidermal thickness increased with increasing concentration of fibrinogen. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (e) Epidermal thickness was calculated as the ratio of the surface area from the basal layer (BL) to the granular cell layer (GL) over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Article Snippet: Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo.

    Techniques: Concentration Assay, Generated, Staining

    Fibrin can be employed as a vehicle to deliver keratinocytes onto dermis. Tissue-engineered skin substitutes were constructed by seeding keratinocytes in fibrin hydrogels containing fibrinogen (4 mg/mL), fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. At the indicated times after cell seeding, tissues were harvested. (a) Paraffin-embedded tissue sections were stained with hematoxylin and eosin. Frozen sections from tissues on day 10 postseeding were immunostained for (b) PCNA (green); or (c) K10 (red), and counterstained with the nuclear dye Hoechst (blue). bar = 50 μm. (d) .

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Fibrin can be employed as a vehicle to deliver keratinocytes onto dermis. Tissue-engineered skin substitutes were constructed by seeding keratinocytes in fibrin hydrogels containing fibrinogen (4 mg/mL), fibronectin (100 ng/mL), and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. At the indicated times after cell seeding, tissues were harvested. (a) Paraffin-embedded tissue sections were stained with hematoxylin and eosin. Frozen sections from tissues on day 10 postseeding were immunostained for (b) PCNA (green); or (c) K10 (red), and counterstained with the nuclear dye Hoechst (blue). bar = 50 μm. (d) .

    Article Snippet: Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo.

    Techniques: Construct, Staining

    Epidermal cell number increased with increasing concentration of fibrin. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL) and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface frozen sections were stained with the nuclear dye Hoechst. The dashed line denotes basement membrane (bar = 50 μm; original magnification 20×). (e) Cell density was measured as the number of cell nuclei per basement membrane length. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: Epidermal cell number increased with increasing concentration of fibrin. (a–d) Bioengineered epidermis was generated by seeding keratinocytes in fibrin hydrogels containing various concentrations of fibrinogen as indicated, fibronectin (100 ng/mL) and KGF (80 ng/mL) on the basement membrane side of decellularized dermis. After 1 week at the air–liquid interface frozen sections were stained with the nuclear dye Hoechst. The dashed line denotes basement membrane (bar = 50 μm; original magnification 20×). (e) Cell density was measured as the number of cell nuclei per basement membrane length. The symbol (*) denotes statistical significance ( p

    Article Snippet: Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo.

    Techniques: Concentration Assay, Generated, Staining

    The thickness of the neoepidermis correlated with the extent of vascularization of the underlying dermal support. In vivo neoepidermis was generated by seeding cultured keratinocytes within fibrin hydrogels onto the prevascularized dermis at 1 week postgrafting. (a) Two weeks after keratinocyte delivery, tissues were excised and paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (b) Epidermal thickness was calculated as the ratio of the surface area from the basal to the granular cell layers over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Journal: Tissue Engineering. Part A

    Article Title: Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

    doi: 10.1089/ten.tea.2010.0125

    Figure Lengend Snippet: The thickness of the neoepidermis correlated with the extent of vascularization of the underlying dermal support. In vivo neoepidermis was generated by seeding cultured keratinocytes within fibrin hydrogels onto the prevascularized dermis at 1 week postgrafting. (a) Two weeks after keratinocyte delivery, tissues were excised and paraffin-embedded tissue sections were stained with hematoxylin and eosin (bar = 50 μm; original magnification 20×). The dashed line denotes basement membrane. (b) Epidermal thickness was calculated as the ratio of the surface area from the basal to the granular cell layers over the length of the basement membrane. The symbol (*) denotes statistical significance ( p

    Article Snippet: Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo.

    Techniques: In Vivo, Generated, Cell Culture, Staining