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  • 99
    Thermo Fisher dispase
    Morphology and function of hepatocytes in subcultures. Hepatocytes were cocultured with 3T3-J2 fibroblasts for 35 days, detached from the substrate by <t>dispase/collagenase</t> treatment, dispersed in a single cell suspension by trypsin digestion, replated,
    Dispase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8835 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dispase ii
    A schematic diagram of Epi-SCs and SKP transplantation for hair follicle regeneration. Dorsal skin tissue in full thickness was collected from neonatal C57 mice ( a ), which was cut into pieces ( b ). The tissue was separated into the epidermis and dermis after treatment with <t>dispase</t> II ( c , d ). Epidermal stem cells (Epi-SCs) were derived from the epidermis as described in the “Methods” section, which grew in monolayer ( e ) and expressed CD49f as detected by immunofluorescence staining (red, g ). Skin-derived precursors (SKPs) were derived from the dermis as described in the “Methods” section, which were grown in spheroids and expressed nestin as detected by immunofluorescence staining (red, h ). Full thickness excisional skin wounds were prepared in nu/nu mice ( i ), and a mixture of Epi-SCs and SKPs in Matrigel was implanted into the wound, which resulted in the growth of black hairs ( j ). Scale bar, 50 μm
    Dispase Ii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1769 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dispase 1
    Cell yield from explants on PET membrane inserts in five different cell culture media. Cells were isolated from PET membrane inserts using <t>dispase</t> and total cell yield per insert was counted. Mann-Whitney U-test was performed (* p
    Dispase 1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor dispase
    Cell yield from explants on PET membrane inserts in five different cell culture media. Cells were isolated from PET membrane inserts using <t>dispase</t> and total cell yield per insert was counted. Mann-Whitney U-test was performed (* p
    Dispase, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson dispase
    naEFCs express mature EC markers and form perfused tubes in vivo. In (A), CFSE-labelled naEFCs mixed with Matrigel prior to injection into the flank of NOD/SCID mice, after 7 days the plugs were removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The upper left image shows the cross section of a CFSE-naEFC generated tube-like structure (green) within which the nuclei of cells can be seen (blue) at 60× mag (arrows). The upper right image is the control plug in which no naEFCs were added. Images represent one experiment of n≥3. Similar sections were stained for CD144 and images captured by confocal microscopy with CFSE-naEFCs (green) exhibiting CD144 (red) as a cross section of a tube (lower left image) and CD144 staining in the junctions of the CFSE-naEFCs (lower right panel). Images are a representative of n≥3. In (B), similar experiments were executed and at day 7 post-implant the mice were injected i.v. with TRITC-lectin prior to exsanguinations, plugs removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The representative image shows the cross section of a CFSE-naEFC generated tube-like structure (green, upper left image), TRITIC-lectin (red, upper right image), DAPI counterstain (blue, lower left image) and the merged image (lower right). In (C), CFSE-naEFCs were digested from explanted Matrigel plugs using <t>dispase</t> and phenotyped for hematopoietic progenitor cell and endothelial cell markers by flow cytometry (right panel); cells from contra-lateral control Matrigel plugs were similarly examined for antigen expression (left panel). In the histograms, the light dotted lines represent unstained cells and the dark lines represent stained cells of a representative of repeated experiments.
    Dispase, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 2540 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Boehringer Mannheim dispase
    PECAM immunofluorescence of ES cell differentiation. <t>Dispase-treated</t> ES cells were plated onto tissue culture dishes on day 0. Representative plates were fixed every other day. Wells were stained with an anti-PECAM antibody and a B-phycoerythrin-conjugated secondary antibody. A, D : day 2; B, E : day 4; C, F : day 6; G, J : day 8; H, K : day 10; I, J : day 12. A–C, G–I : fluorescence photomicrograph; D–F, J–L : the corresponding phase contrast photomicrograph. Arrowheads in H point to PECAM strain at cell borders. Scale bar, 30 μm.
    Dispase, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Corning Life Sciences dispase
    Evaluation of the cytotoxicity induced by MWCNTs on HBMECs grown on plastic or matrigel HBMECs were plated on standard tissue culture plastic or matrigel coated plastic and recovered overnight. HBMECs grown on plastic (A) or Matrigel (B) were treated for 24 h with increasing doses of different types of MWCNTs as indicated. Cells were lysed, pelleted, and the supernatant was analyzed for ATP content as a measure of cell viability using the CellTiter-Glo assay. Three independent biological replicates were performed with at least three technical replicates per treatment group. Endpoint analysis shows the groups that are significantly different (*p≤0.05, **p≤0.01) at the highest dose (ANOVA; post-hoc T-Test). In parallel, lysates of similarly treated cells grown on plastic (C) or Matrigel (D) were collected and probed for cleaved PARP, a protein indicative of apoptosis, via western blot analysis. GAPDH was used as a loading control. Lysates from U87 cells treated with 10 μM doxorubicin was used as a positive control. Cells grown on matrigel were isolated using <t>dispase</t> to degrade the matrigel without affecting HBMEC cell integrity. The graph below the western blot displays average fold change of cleaved PARP protein levels relative to GAPDH loading control for three independent biological replicates. Fold change is displayed as the mean ± standard deviation of each experiment.
    Dispase, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 485 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    FUJIFILM dispase
    Primary culture procedure. (a) Disinfection and washing of biopsied oral mucosal tissue. (b) Photograph of washed oral mucosal tissue in 35 mm dish. (c) Cutting of oral tissue into 5 mm cubes for <t>dispase</t> treatment. (d) Separation of epithelial layer from connective tissue. (e) Cutting of separated epithelial tissue into 1 mm cubes for trypsin–EDTA treatment. (f) 1000 μL micropipette tip with cutting down the tip at 0-mm long (Normal), 1-mm long, and 2-mm long. (g) Counting suspended cells. Seeding cells into thermo-responsive cell culture inserts at a density of 8 × 10 4 viable cells/cm 2 .
    Dispase, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche dispase ii
    The dermis basement membrane obtained by the three different techniques stained with Collagen IV MoAb. (a) <t>Dispase</t> II, (b) Trypsin, (c) the manual method. Only the manual method left an integral structure. Arrows indicate the absence of basement membrane. Original magnification: 10x. AAGD: alloplastic acellular glycerolized dermis; BM: basement membrane.
    Dispase Ii, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 6989 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Sanko Junyaku Co Ltd dispase ii
    The dermis basement membrane obtained by the three different techniques stained with Collagen IV MoAb. (a) <t>Dispase</t> II, (b) Trypsin, (c) the manual method. Only the manual method left an integral structure. Arrows indicate the absence of basement membrane. Original magnification: 10x. AAGD: alloplastic acellular glycerolized dermis; BM: basement membrane.
    Dispase Ii, supplied by Sanko Junyaku Co Ltd, used in various techniques. Bioz Stars score: 93/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    STEMCELL Technologies Inc dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 94/100, based on 1837 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Atlanta Biologicals dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by Atlanta Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Fisher Scientific dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Valiant dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by Valiant, used in various techniques. Bioz Stars score: 90/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Roche of dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Of Dispase, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Scientific Laboratory Supplies Ltd dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by Scientific Laboratory Supplies Ltd, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dispase/product/Scientific Laboratory Supplies Ltd
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    90
    Dojindo Labs dispase
    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a <t>dispase</t> digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.
    Dispase, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Morphology and function of hepatocytes in subcultures. Hepatocytes were cocultured with 3T3-J2 fibroblasts for 35 days, detached from the substrate by dispase/collagenase treatment, dispersed in a single cell suspension by trypsin digestion, replated,

    Journal: Biotechnology and bioengineering

    Article Title: A New Technique for Primary Hepatocyte Expansion In Vitro

    doi: 10.1002/bit.21911

    Figure Lengend Snippet: Morphology and function of hepatocytes in subcultures. Hepatocytes were cocultured with 3T3-J2 fibroblasts for 35 days, detached from the substrate by dispase/collagenase treatment, dispersed in a single cell suspension by trypsin digestion, replated,

    Article Snippet: Long-term cultures of hepatocytes (35 days after seeding) were treated with collagenase (1 mg/mL; Sigma Chemical Co.) and dispase (1 mg/mL; Gibco).

    Techniques:

    A schematic diagram of Epi-SCs and SKP transplantation for hair follicle regeneration. Dorsal skin tissue in full thickness was collected from neonatal C57 mice ( a ), which was cut into pieces ( b ). The tissue was separated into the epidermis and dermis after treatment with dispase II ( c , d ). Epidermal stem cells (Epi-SCs) were derived from the epidermis as described in the “Methods” section, which grew in monolayer ( e ) and expressed CD49f as detected by immunofluorescence staining (red, g ). Skin-derived precursors (SKPs) were derived from the dermis as described in the “Methods” section, which were grown in spheroids and expressed nestin as detected by immunofluorescence staining (red, h ). Full thickness excisional skin wounds were prepared in nu/nu mice ( i ), and a mixture of Epi-SCs and SKPs in Matrigel was implanted into the wound, which resulted in the growth of black hairs ( j ). Scale bar, 50 μm

    Journal: Stem Cell Research & Therapy

    Article Title: PI3K/Akt signaling pathway is essential for de novo hair follicle regeneration

    doi: 10.1186/s13287-020-01650-6

    Figure Lengend Snippet: A schematic diagram of Epi-SCs and SKP transplantation for hair follicle regeneration. Dorsal skin tissue in full thickness was collected from neonatal C57 mice ( a ), which was cut into pieces ( b ). The tissue was separated into the epidermis and dermis after treatment with dispase II ( c , d ). Epidermal stem cells (Epi-SCs) were derived from the epidermis as described in the “Methods” section, which grew in monolayer ( e ) and expressed CD49f as detected by immunofluorescence staining (red, g ). Skin-derived precursors (SKPs) were derived from the dermis as described in the “Methods” section, which were grown in spheroids and expressed nestin as detected by immunofluorescence staining (red, h ). Full thickness excisional skin wounds were prepared in nu/nu mice ( i ), and a mixture of Epi-SCs and SKPs in Matrigel was implanted into the wound, which resulted in the growth of black hairs ( j ). Scale bar, 50 μm

    Article Snippet: The tissue was washed 3 times in PBS, cut into 2~3 mm2 pieces, and digested with 0.35% Dispase II (sigma) for 40 min at 37 °C.

    Techniques: Transplantation Assay, Mouse Assay, Derivative Assay, Immunofluorescence, Staining

    Cell yield from explants on PET membrane inserts in five different cell culture media. Cells were isolated from PET membrane inserts using dispase and total cell yield per insert was counted. Mann-Whitney U-test was performed (* p

    Journal: Cytotechnology

    Article Title: Comparison of culture media indicates a role for autologous serum in enhancing phenotypic preservation of rabbit limbal stem cells in explant culture

    doi: 10.1007/s10616-017-0171-7

    Figure Lengend Snippet: Cell yield from explants on PET membrane inserts in five different cell culture media. Cells were isolated from PET membrane inserts using dispase and total cell yield per insert was counted. Mann-Whitney U-test was performed (* p

    Article Snippet: After dispase II digestion, epithelial sheets detached from PET inserts were incubated with 0.25% trypsin-EDTA (Sigma) solution at 37 °C for ten min., with shaking at 80 rpm.

    Techniques: Positron Emission Tomography, Cell Culture, Isolation, MANN-WHITNEY

    Targeting of Dsg3 leads to p38 MAPK-dependent loss of cell cohesion. A , targeting of Dsg3 function by either incubation with AK23 or siRNA-mediated depletion of Dsg3 protein levels in HaCaT cells resulted in increased cell monolayer fragmentation in dispase-based

    Journal: The Journal of Biological Chemistry

    Article Title: Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

    doi: 10.1074/jbc.M113.489336

    Figure Lengend Snippet: Targeting of Dsg3 leads to p38 MAPK-dependent loss of cell cohesion. A , targeting of Dsg3 function by either incubation with AK23 or siRNA-mediated depletion of Dsg3 protein levels in HaCaT cells resulted in increased cell monolayer fragmentation in dispase-based

    Article Snippet: After removing the medium and washing with PBS, 150 μl of the enzyme dispase ( > 2.4 units/ml Dispase II in Hanks' buffered saline solution (Sigma)) were incubated for 20 min at 37 °C to detach the whole cell monolayer from the well bottom.

    Techniques: Incubation

    naEFCs express mature EC markers and form perfused tubes in vivo. In (A), CFSE-labelled naEFCs mixed with Matrigel prior to injection into the flank of NOD/SCID mice, after 7 days the plugs were removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The upper left image shows the cross section of a CFSE-naEFC generated tube-like structure (green) within which the nuclei of cells can be seen (blue) at 60× mag (arrows). The upper right image is the control plug in which no naEFCs were added. Images represent one experiment of n≥3. Similar sections were stained for CD144 and images captured by confocal microscopy with CFSE-naEFCs (green) exhibiting CD144 (red) as a cross section of a tube (lower left image) and CD144 staining in the junctions of the CFSE-naEFCs (lower right panel). Images are a representative of n≥3. In (B), similar experiments were executed and at day 7 post-implant the mice were injected i.v. with TRITC-lectin prior to exsanguinations, plugs removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The representative image shows the cross section of a CFSE-naEFC generated tube-like structure (green, upper left image), TRITIC-lectin (red, upper right image), DAPI counterstain (blue, lower left image) and the merged image (lower right). In (C), CFSE-naEFCs were digested from explanted Matrigel plugs using dispase and phenotyped for hematopoietic progenitor cell and endothelial cell markers by flow cytometry (right panel); cells from contra-lateral control Matrigel plugs were similarly examined for antigen expression (left panel). In the histograms, the light dotted lines represent unstained cells and the dark lines represent stained cells of a representative of repeated experiments.

    Journal: PLoS ONE

    Article Title: Characterization of a Distinct Population of Circulating Human Non-Adherent Endothelial Forming Cells and Their Recruitment via Intercellular Adhesion Molecule-3

    doi: 10.1371/journal.pone.0046996

    Figure Lengend Snippet: naEFCs express mature EC markers and form perfused tubes in vivo. In (A), CFSE-labelled naEFCs mixed with Matrigel prior to injection into the flank of NOD/SCID mice, after 7 days the plugs were removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The upper left image shows the cross section of a CFSE-naEFC generated tube-like structure (green) within which the nuclei of cells can be seen (blue) at 60× mag (arrows). The upper right image is the control plug in which no naEFCs were added. Images represent one experiment of n≥3. Similar sections were stained for CD144 and images captured by confocal microscopy with CFSE-naEFCs (green) exhibiting CD144 (red) as a cross section of a tube (lower left image) and CD144 staining in the junctions of the CFSE-naEFCs (lower right panel). Images are a representative of n≥3. In (B), similar experiments were executed and at day 7 post-implant the mice were injected i.v. with TRITC-lectin prior to exsanguinations, plugs removed, processed and sections counterstained for nuclei with DAPI prior to imaging by confocal microscopy. The representative image shows the cross section of a CFSE-naEFC generated tube-like structure (green, upper left image), TRITIC-lectin (red, upper right image), DAPI counterstain (blue, lower left image) and the merged image (lower right). In (C), CFSE-naEFCs were digested from explanted Matrigel plugs using dispase and phenotyped for hematopoietic progenitor cell and endothelial cell markers by flow cytometry (right panel); cells from contra-lateral control Matrigel plugs were similarly examined for antigen expression (left panel). In the histograms, the light dotted lines represent unstained cells and the dark lines represent stained cells of a representative of repeated experiments.

    Article Snippet: Plugs were macerated and incubated in 3 ml of Dispase (BD Biosciences) for 2 hours at RT to isolate the cells within the excised plug.

    Techniques: In Vivo, Injection, Mouse Assay, Imaging, Confocal Microscopy, Generated, Staining, Flow Cytometry, Cytometry, Expressing

    PECAM immunofluorescence of ES cell differentiation. Dispase-treated ES cells were plated onto tissue culture dishes on day 0. Representative plates were fixed every other day. Wells were stained with an anti-PECAM antibody and a B-phycoerythrin-conjugated secondary antibody. A, D : day 2; B, E : day 4; C, F : day 6; G, J : day 8; H, K : day 10; I, J : day 12. A–C, G–I : fluorescence photomicrograph; D–F, J–L : the corresponding phase contrast photomicrograph. Arrowheads in H point to PECAM strain at cell borders. Scale bar, 30 μm.

    Journal: The American Journal of Pathology

    Article Title: Developmental Platelet Endothelial Cell Adhesion Molecule Expression Suggests Multiple Roles for a Vascular Adhesion Molecule

    doi:

    Figure Lengend Snippet: PECAM immunofluorescence of ES cell differentiation. Dispase-treated ES cells were plated onto tissue culture dishes on day 0. Representative plates were fixed every other day. Wells were stained with an anti-PECAM antibody and a B-phycoerythrin-conjugated secondary antibody. A, D : day 2; B, E : day 4; C, F : day 6; G, J : day 8; H, K : day 10; I, J : day 12. A–C, G–I : fluorescence photomicrograph; D–F, J–L : the corresponding phase contrast photomicrograph. Arrowheads in H point to PECAM strain at cell borders. Scale bar, 30 μm.

    Article Snippet: These cultures were then dissociated into clumps by treatment with Dispase (Boehringer Mannheim, Indianapolis, IN) on day 0.

    Techniques: Immunofluorescence, Cell Differentiation, Staining, Fluorescence

    Double immunofluorescence for PECAM and CD34 in ES cells and during ES cell differentiation. A, F : undifferentiated ES cells, fixed 3 days after passage. B–E, G–J : Dispase-treated ES cells were cultured in Petri dishes for 3 days, then plated onto tissue culture dishes. Representative plates were fixed every day, beginning with day 5. Wells were stained with biotin-conjugated Mec 13.3 ( A–C, E ) and an anti-CD34 rat monoclonal antibody ( F–I ). Control plates had either the anti-PECAM primary antibody ( D ) or the anti-CD34 antibody primary antibody ( J ) omitted. Detection was with streptavidin-conjugated B-phycoerythrin and donkey anti-rat FITC. A, F , undifferentiated ES cells; B, G , day 6; C–E, H–J , day 10. A–E , PECAM-specific B-phycoerythrin fluorescence; F–J , CD34-specific FITC fluorescence of the same wells. In B and G , the arrow shows cells that are CD34-negative and PECAM-positive and the arrowhead indicates cells that are double positive.

    Journal: The American Journal of Pathology

    Article Title: Developmental Platelet Endothelial Cell Adhesion Molecule Expression Suggests Multiple Roles for a Vascular Adhesion Molecule

    doi:

    Figure Lengend Snippet: Double immunofluorescence for PECAM and CD34 in ES cells and during ES cell differentiation. A, F : undifferentiated ES cells, fixed 3 days after passage. B–E, G–J : Dispase-treated ES cells were cultured in Petri dishes for 3 days, then plated onto tissue culture dishes. Representative plates were fixed every day, beginning with day 5. Wells were stained with biotin-conjugated Mec 13.3 ( A–C, E ) and an anti-CD34 rat monoclonal antibody ( F–I ). Control plates had either the anti-PECAM primary antibody ( D ) or the anti-CD34 antibody primary antibody ( J ) omitted. Detection was with streptavidin-conjugated B-phycoerythrin and donkey anti-rat FITC. A, F , undifferentiated ES cells; B, G , day 6; C–E, H–J , day 10. A–E , PECAM-specific B-phycoerythrin fluorescence; F–J , CD34-specific FITC fluorescence of the same wells. In B and G , the arrow shows cells that are CD34-negative and PECAM-positive and the arrowhead indicates cells that are double positive.

    Article Snippet: These cultures were then dissociated into clumps by treatment with Dispase (Boehringer Mannheim, Indianapolis, IN) on day 0.

    Techniques: Immunofluorescence, Cell Differentiation, Cell Culture, Staining, Fluorescence

    Evaluation of the cytotoxicity induced by MWCNTs on HBMECs grown on plastic or matrigel HBMECs were plated on standard tissue culture plastic or matrigel coated plastic and recovered overnight. HBMECs grown on plastic (A) or Matrigel (B) were treated for 24 h with increasing doses of different types of MWCNTs as indicated. Cells were lysed, pelleted, and the supernatant was analyzed for ATP content as a measure of cell viability using the CellTiter-Glo assay. Three independent biological replicates were performed with at least three technical replicates per treatment group. Endpoint analysis shows the groups that are significantly different (*p≤0.05, **p≤0.01) at the highest dose (ANOVA; post-hoc T-Test). In parallel, lysates of similarly treated cells grown on plastic (C) or Matrigel (D) were collected and probed for cleaved PARP, a protein indicative of apoptosis, via western blot analysis. GAPDH was used as a loading control. Lysates from U87 cells treated with 10 μM doxorubicin was used as a positive control. Cells grown on matrigel were isolated using dispase to degrade the matrigel without affecting HBMEC cell integrity. The graph below the western blot displays average fold change of cleaved PARP protein levels relative to GAPDH loading control for three independent biological replicates. Fold change is displayed as the mean ± standard deviation of each experiment.

    Journal: Toxicology in vitro : an international journal published in association with BIBRA

    Article Title: Evaluation of multiwalled carbon nanotube cytotoxicity in cultures of human brain microvascular endothelial cells grown on plastic or basement membrane

    doi: 10.1016/j.tiv.2017.03.002

    Figure Lengend Snippet: Evaluation of the cytotoxicity induced by MWCNTs on HBMECs grown on plastic or matrigel HBMECs were plated on standard tissue culture plastic or matrigel coated plastic and recovered overnight. HBMECs grown on plastic (A) or Matrigel (B) were treated for 24 h with increasing doses of different types of MWCNTs as indicated. Cells were lysed, pelleted, and the supernatant was analyzed for ATP content as a measure of cell viability using the CellTiter-Glo assay. Three independent biological replicates were performed with at least three technical replicates per treatment group. Endpoint analysis shows the groups that are significantly different (*p≤0.05, **p≤0.01) at the highest dose (ANOVA; post-hoc T-Test). In parallel, lysates of similarly treated cells grown on plastic (C) or Matrigel (D) were collected and probed for cleaved PARP, a protein indicative of apoptosis, via western blot analysis. GAPDH was used as a loading control. Lysates from U87 cells treated with 10 μM doxorubicin was used as a positive control. Cells grown on matrigel were isolated using dispase to degrade the matrigel without affecting HBMEC cell integrity. The graph below the western blot displays average fold change of cleaved PARP protein levels relative to GAPDH loading control for three independent biological replicates. Fold change is displayed as the mean ± standard deviation of each experiment.

    Article Snippet: For HBMECs grown on Matrigel®, cells were isolated by aspirating normal growth media and adding equivalent culture volumes of Dispase (Corning), incubating at 37 °C for 30 min, then washing 2x with ice cold 1xPBS prior to lysis.

    Techniques: Glo Assay, Western Blot, Positive Control, Isolation, Standard Deviation

    Primary culture procedure. (a) Disinfection and washing of biopsied oral mucosal tissue. (b) Photograph of washed oral mucosal tissue in 35 mm dish. (c) Cutting of oral tissue into 5 mm cubes for dispase treatment. (d) Separation of epithelial layer from connective tissue. (e) Cutting of separated epithelial tissue into 1 mm cubes for trypsin–EDTA treatment. (f) 1000 μL micropipette tip with cutting down the tip at 0-mm long (Normal), 1-mm long, and 2-mm long. (g) Counting suspended cells. Seeding cells into thermo-responsive cell culture inserts at a density of 8 × 10 4 viable cells/cm 2 .

    Journal: Regenerative Therapy

    Article Title: A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application

    doi: 10.1016/j.reth.2019.11.007

    Figure Lengend Snippet: Primary culture procedure. (a) Disinfection and washing of biopsied oral mucosal tissue. (b) Photograph of washed oral mucosal tissue in 35 mm dish. (c) Cutting of oral tissue into 5 mm cubes for dispase treatment. (d) Separation of epithelial layer from connective tissue. (e) Cutting of separated epithelial tissue into 1 mm cubes for trypsin–EDTA treatment. (f) 1000 μL micropipette tip with cutting down the tip at 0-mm long (Normal), 1-mm long, and 2-mm long. (g) Counting suspended cells. Seeding cells into thermo-responsive cell culture inserts at a density of 8 × 10 4 viable cells/cm 2 .

    Article Snippet: Next, each tissue was cut into 5-mm cubes for effective enzymatic reactions ( c); 1000 U/mL dispase (Wako, Osaka, Japan) was reacted with the tissue in a new 35-mm cell culture dish in a CO2 -atmosphere controlled incubator at 37 °C for 2 h. After the incubation, the epithelial layers were separated from the connective tissue using two pairs of tweezers and then placed in fresh DMEM-AB ( d).

    Techniques: Cell Culture

    The dermis basement membrane obtained by the three different techniques stained with Collagen IV MoAb. (a) Dispase II, (b) Trypsin, (c) the manual method. Only the manual method left an integral structure. Arrows indicate the absence of basement membrane. Original magnification: 10x. AAGD: alloplastic acellular glycerolized dermis; BM: basement membrane.

    Journal: Journal of Biomedicine and Biotechnology

    Article Title: Preparation and Characterization of a Novel Skin Substitute

    doi: 10.1155/2010/840363

    Figure Lengend Snippet: The dermis basement membrane obtained by the three different techniques stained with Collagen IV MoAb. (a) Dispase II, (b) Trypsin, (c) the manual method. Only the manual method left an integral structure. Arrows indicate the absence of basement membrane. Original magnification: 10x. AAGD: alloplastic acellular glycerolized dermis; BM: basement membrane.

    Article Snippet: The first method used dispase II, which cleaves the anchoring system between the dermis and epidermis.

    Techniques: Staining

    Histomorphological analysis of the dermis obtained by the three different techniques nebulized with keratinocytes. The sample refers to the 14th day ((a), (b), and (c)) and to the 21st day ((d), (e), and (f)), (a) and (d) Dispase II, (b) and (e) Trypsin, (c) and (f): the manual method. Nebulized cell adherence was observed only in the one prepared manually. Original magnification: 10x ((a), (d), (e) and (f)) 5x ((b) and (c)). AAGD: alloplastic acellular glycerolized dermis; NFE: newly formed epithelium.

    Journal: Journal of Biomedicine and Biotechnology

    Article Title: Preparation and Characterization of a Novel Skin Substitute

    doi: 10.1155/2010/840363

    Figure Lengend Snippet: Histomorphological analysis of the dermis obtained by the three different techniques nebulized with keratinocytes. The sample refers to the 14th day ((a), (b), and (c)) and to the 21st day ((d), (e), and (f)), (a) and (d) Dispase II, (b) and (e) Trypsin, (c) and (f): the manual method. Nebulized cell adherence was observed only in the one prepared manually. Original magnification: 10x ((a), (d), (e) and (f)) 5x ((b) and (c)). AAGD: alloplastic acellular glycerolized dermis; NFE: newly formed epithelium.

    Article Snippet: The first method used dispase II, which cleaves the anchoring system between the dermis and epidermis.

    Techniques:

    Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a dispase digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: A Method for Cryogenic Preservation of Human Biopsy Specimens and Subsequent Organoid Culture

    doi: 10.1016/j.jcmgh.2018.04.008

    Figure Lengend Snippet: Schematic of the methods developed that allow successful generation of patient biopsy-derived epithelial organoids after cryopreservation. ( A ) The process of endoscopic biopsy collection followed by cryopreservation in a simple freezing medium can be accomplished in typical clinical settings with readily available equipment. After cryopreservation, organoid cultures can be established using 3 different techniques. Technique 1 ( left ) used a dispase digestion to isolate crypts from freshly thawed tissue. Technique 2 ( middle–left ) adds a step relative to technique 1 in which the entire biopsy specimen is embedded in Matrigel and allowed to repair from the freezing process before tissue digestion and crypt isolation. Technique 3 ( middle–right ) is very similar to technique 1 but uses a gentle EDTA treatment to separate the epithelium from the mesenchyme. Technique 4 ( right ) involves isolation of epithelial crypts before cyropreservation so that, upon thawing, cultures can be seeded immediately without additional tissue manipulations. All 4 techniques result in pure organoid cultures 2 weeks after initially thawing the biopsy specimen. ( B ) Organoids lines were generated using techniques 1 and 2. ( C ) Organoid lines were generated using technique 3. ( D ) Organoid lines were generated using technique 4.

    Article Snippet: To separate the epithelium from the underlying cell layers, minced biopsy specimens then were incubated in dispase (07923; STEMCELL Technologies, Vancouver, Canada) for 30 minutes on ice.

    Techniques: Derivative Assay, Isolation, Generated