dispase ii  (Millipore)


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    Name:
    Dispase II
    Description:

    Catalog Number:
    d4693
    Price:
    None
    Applications:
    The enzyme from Sigma has been used in developing a protocol for ex vivo culture of mouse embryonic mammary buds. It has been used in the treatment of rat heart pieces during the isolation of mitochondria from rat heart. It has also been used for the isolation of dental pulp stem cells (DPSCs) by enzymatic hydrolysis. These cells have been compared with DPSCs isolated by explant method to analyse their stem cell and differentiation properties.Dispase II has been used for Fluorescence-Activated Cell Sorting (FACS). It has also been used for separating visceral yolk sac layers.
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    Structured Review

    Millipore dispase ii
    Tissue viability analysis. (A) Viability index of the epidermal sheet determined by CCK-8 assay. There is no significant difference in the viability index between the 8- and 10-h digestion groups, while the value of the 12-h digestion group is significantly lower than that of the 8-h digestion group. n = 5. (B) The percentage of viable cells detected by Hoe/PI staining. With the duration of <t>Dispase</t> II digestion prolonging, the percentage of viable cells in the epidermal sheet decreases gradually. n = 5.

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    Images

    1) Product Images from "Evaluation of Dermal Substitute in a Novel Co-Transplantation Model with Autologous Epidermal Sheet"

    Article Title: Evaluation of Dermal Substitute in a Novel Co-Transplantation Model with Autologous Epidermal Sheet

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0049448

    Tissue viability analysis. (A) Viability index of the epidermal sheet determined by CCK-8 assay. There is no significant difference in the viability index between the 8- and 10-h digestion groups, while the value of the 12-h digestion group is significantly lower than that of the 8-h digestion group. n = 5. (B) The percentage of viable cells detected by Hoe/PI staining. With the duration of Dispase II digestion prolonging, the percentage of viable cells in the epidermal sheet decreases gradually. n = 5.
    Figure Legend Snippet: Tissue viability analysis. (A) Viability index of the epidermal sheet determined by CCK-8 assay. There is no significant difference in the viability index between the 8- and 10-h digestion groups, while the value of the 12-h digestion group is significantly lower than that of the 8-h digestion group. n = 5. (B) The percentage of viable cells detected by Hoe/PI staining. With the duration of Dispase II digestion prolonging, the percentage of viable cells in the epidermal sheet decreases gradually. n = 5.

    Techniques Used: CCK-8 Assay, Staining

    2) Product Images from "PI3K/Akt signaling pathway is essential for de novo hair follicle regeneration"

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

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-020-01650-6

    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
    Figure Legend 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

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

    3) Product Images from "Deposition of microparticles by neutrophils onto inflamed epithelium: a new mechanism to disrupt epithelial intercellular adhesions and promote transepithelial migration"

    Article Title: Deposition of microparticles by neutrophils onto inflamed epithelium: a new mechanism to disrupt epithelial intercellular adhesions and promote transepithelial migration

    Journal: The FASEB Journal

    doi: 10.1096/fj.201600734R

    PMN-MP binding to IECs results in MMP-9 dependent disruption of epithelial cell-to-cell adhesions. Dispase-based dissociation assay was used to assess the strength of intercellular adhesions and monolayer integrity after the indicated treatments. A ) Freshly isolated PMNs were added to IEC monolayers and induced to adhere in the presence of 100 nM fMLF (2 × 10 6 PMNs/well, 3 h, 37°C). PMN adhesion to IECs resulted in significant fragmentation of IEC monolayers, which was dependent on MMP-9 activity, after Dispase treatment. *** P
    Figure Legend Snippet: PMN-MP binding to IECs results in MMP-9 dependent disruption of epithelial cell-to-cell adhesions. Dispase-based dissociation assay was used to assess the strength of intercellular adhesions and monolayer integrity after the indicated treatments. A ) Freshly isolated PMNs were added to IEC monolayers and induced to adhere in the presence of 100 nM fMLF (2 × 10 6 PMNs/well, 3 h, 37°C). PMN adhesion to IECs resulted in significant fragmentation of IEC monolayers, which was dependent on MMP-9 activity, after Dispase treatment. *** P

    Techniques Used: Binding Assay, Isolation, Activity Assay

    4) Product Images from "Arrhythmogenic cardiomyopathy related DSG2 mutations affect desmosomal cadherin binding kinetics"

    Article Title: Arrhythmogenic cardiomyopathy related DSG2 mutations affect desmosomal cadherin binding kinetics

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-13737-x

    Confluent monolayers of transfected cells were subjected to dispase assay. Decreased number of fragments indicates relatively enhanced cell-cell adhesion. Box and whiskers plots represent the number of fragments in six independent experiments for Dsg2 wildtype, p.D105E (p.D154E; \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$p
    Figure Legend Snippet: Confluent monolayers of transfected cells were subjected to dispase assay. Decreased number of fragments indicates relatively enhanced cell-cell adhesion. Box and whiskers plots represent the number of fragments in six independent experiments for Dsg2 wildtype, p.D105E (p.D154E; \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$p

    Techniques Used: Transfection

    ( A ) Crystal structure of Dsg2 homodimer shown in ribbon representation [ 12 , pdbID: 5ERD]. Coordinated Ca 2+ are shown as blue spheres. ( B ) Schematic of cantilever tip and sample substrate functionalized with Dsg2 for AFM single molecule force spectroscopy. Both, cantilever and substrate are modified with covalently immobilized Dsg2 fragments. ( C ) SMFS working principle and typical force-distance cycle exposing a specific dissociation event. The striated area under the curve represents the dissipated work to break a Dsg2 homodimer. ( D ) Principle of dispase assay for measuring cell-cell adhesion. Cells grown to confluency (a) are lifted up by dispase treatment and exposed to mechanical stress by shaking of cell cultures. Resulting islets of the monolayer are counted and recombinant cell cultures of mutant and wildtype are compared (b).
    Figure Legend Snippet: ( A ) Crystal structure of Dsg2 homodimer shown in ribbon representation [ 12 , pdbID: 5ERD]. Coordinated Ca 2+ are shown as blue spheres. ( B ) Schematic of cantilever tip and sample substrate functionalized with Dsg2 for AFM single molecule force spectroscopy. Both, cantilever and substrate are modified with covalently immobilized Dsg2 fragments. ( C ) SMFS working principle and typical force-distance cycle exposing a specific dissociation event. The striated area under the curve represents the dissipated work to break a Dsg2 homodimer. ( D ) Principle of dispase assay for measuring cell-cell adhesion. Cells grown to confluency (a) are lifted up by dispase treatment and exposed to mechanical stress by shaking of cell cultures. Resulting islets of the monolayer are counted and recombinant cell cultures of mutant and wildtype are compared (b).

    Techniques Used: Spectroscopy, Modification, Recombinant, Mutagenesis

    5) Product Images from "Pemphigus Vulgaris IgG Cause Loss of Desmoglein-Mediated Adhesion and Keratinocyte Dissociation Independent of Epidermal Growth Factor Receptor"

    Article Title: Pemphigus Vulgaris IgG Cause Loss of Desmoglein-Mediated Adhesion and Keratinocyte Dissociation Independent of Epidermal Growth Factor Receptor

    Journal: The American Journal of Pathology

    doi: 10.2353/ajpath.2009.080392

    EGF but not PV-IgG caused keratinocyte dissociation dependent on EGFR and c-Src signaling. Keratinocyte dissociation was further quantified using a dispase-base assay. EGF treatment resulted in a strong increase of cell fragments compared with controls. This increase was partially blocked by simultaneous incubation with EGFR kinase inhibitor GW2974 or c-Src inhibitor PP2. In contrast, PV-IgG-mediated acantholysis was independent of EGFR kinase or c-Src because GW2974 and PP2 did not prevent increase in cell fragments. However, cotreatment with CNF1 and CNFy inhibited PV-IgG 4-induced cell dissociation. Similarly, in NHEK cells GW2974 and PP2 application were only sufficient to inhibit EGF- but not PV-IgG-mediated cell layer fragmentation ( n = 6 for each condition).
    Figure Legend Snippet: EGF but not PV-IgG caused keratinocyte dissociation dependent on EGFR and c-Src signaling. Keratinocyte dissociation was further quantified using a dispase-base assay. EGF treatment resulted in a strong increase of cell fragments compared with controls. This increase was partially blocked by simultaneous incubation with EGFR kinase inhibitor GW2974 or c-Src inhibitor PP2. In contrast, PV-IgG-mediated acantholysis was independent of EGFR kinase or c-Src because GW2974 and PP2 did not prevent increase in cell fragments. However, cotreatment with CNF1 and CNFy inhibited PV-IgG 4-induced cell dissociation. Similarly, in NHEK cells GW2974 and PP2 application were only sufficient to inhibit EGF- but not PV-IgG-mediated cell layer fragmentation ( n = 6 for each condition).

    Techniques Used: Incubation

    6) Product Images from "Dsg2 Upregulation as a Rescue Mechanism in Pemphigus"

    Article Title: Dsg2 Upregulation as a Rescue Mechanism in Pemphigus

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.581370

    Dsg2 is upregulated in murine Dsg3 ko keratinocytes. (A) Dispase-based keratinocyte dissociation assay of stable Dsg3 wt and ko keratinocytes showing compromised intercellular adhesion in cells lacking Dsg3. Columns indicate mean value ± SEM, *P
    Figure Legend Snippet: Dsg2 is upregulated in murine Dsg3 ko keratinocytes. (A) Dispase-based keratinocyte dissociation assay of stable Dsg3 wt and ko keratinocytes showing compromised intercellular adhesion in cells lacking Dsg3. Columns indicate mean value ± SEM, *P

    Techniques Used:

    Heterophilic Dsg2-Dsg3 interactions were less susceptible to autoantibody-induced direct inhibition of Dsg interaction. Binding frequencies of cell-free AFM measurements probing Dsg3-Dsg3 and Dsg2-Dsg3 interaction pairs with and without (ctr) incubation of either a monoclonal anti-Dsg3 antibody (AK23) (A) or PV3-IgG (B) for 1 h (n ≥ 5, each 2500 force-distance curves). Heterophilic Dsg2-Dsg3 interactions are less susceptible against direct inhibition by pathogenic Dsg3 antibodies than homophilic Dsg3 interactions. (C) Dispase-based keratinocyte dissociation assay of Dsg3 wt and ko keratinocytes after treatment with control (ctr) IgG or AK23 for 24 h showing loss of intercellular adhesion after AK23 in wt cells only (n = 3). (D) Keratinocyte dissociation assay of Dsg3 ko keratinocytes after treatment with ctr-IgG, AK23 or an inhibitory anti-Dsg2 antibody (aDsg2) for 24 h revealing drastic effect of aDsg2 antibody in cells lacking Dsg3 (n = 4). Columns indicate mean value normalized to ctr ± SEM, *P
    Figure Legend Snippet: Heterophilic Dsg2-Dsg3 interactions were less susceptible to autoantibody-induced direct inhibition of Dsg interaction. Binding frequencies of cell-free AFM measurements probing Dsg3-Dsg3 and Dsg2-Dsg3 interaction pairs with and without (ctr) incubation of either a monoclonal anti-Dsg3 antibody (AK23) (A) or PV3-IgG (B) for 1 h (n ≥ 5, each 2500 force-distance curves). Heterophilic Dsg2-Dsg3 interactions are less susceptible against direct inhibition by pathogenic Dsg3 antibodies than homophilic Dsg3 interactions. (C) Dispase-based keratinocyte dissociation assay of Dsg3 wt and ko keratinocytes after treatment with control (ctr) IgG or AK23 for 24 h showing loss of intercellular adhesion after AK23 in wt cells only (n = 3). (D) Keratinocyte dissociation assay of Dsg3 ko keratinocytes after treatment with ctr-IgG, AK23 or an inhibitory anti-Dsg2 antibody (aDsg2) for 24 h revealing drastic effect of aDsg2 antibody in cells lacking Dsg3 (n = 4). Columns indicate mean value normalized to ctr ± SEM, *P

    Techniques Used: Inhibition, Binding Assay, Incubation

    7) Product Images from "OM-101 Decreases the Fibrotic Response Associated with Proliferative Vitreoretinopathy"

    Article Title: OM-101 Decreases the Fibrotic Response Associated with Proliferative Vitreoretinopathy

    Journal: Journal of Ophthalmology

    doi: 10.1155/2017/1606854

    Funduscopy and OCT in mice eyes after dispase injection. Mice injected with Hepes only (control) demonstrated normal retina (a). Mice injected with dispase demonstrating RD with fibrotic tissue in front of the retina (b). Normal retina on OCT (c) and detached retina with loss of normal appearance of the retina (d).
    Figure Legend Snippet: Funduscopy and OCT in mice eyes after dispase injection. Mice injected with Hepes only (control) demonstrated normal retina (a). Mice injected with dispase demonstrating RD with fibrotic tissue in front of the retina (b). Normal retina on OCT (c) and detached retina with loss of normal appearance of the retina (d).

    Techniques Used: Mouse Assay, Injection

    Histological staining (H E) of induced RD and PVR in retina treated with OM-101. (a) Histological staining (H E) demonstrated the following: left (control/Hepes), normal architecture; middle (dispase + PBS), abnormal morphology with inflammatory cells, fibroblasts, and pigment stain RPE cells through the retina denoting PVR; and right (dispase + OM 101), OM 101 treatment maintained the normal structure of the retina. Scale bar 100 μ m or 200 μ m. (b) Number of mice developing RD or PVR. (c) Severity score of the mice retina measured by thickness of the RD, number of inflammatory cells, loss of normal structure, and the appearance of RPE cells inside the retina ( N = 25). Statistics were computed using Student's t -test (two-tailed distribution equal variance). Data is expressed as the mean ± SD.
    Figure Legend Snippet: Histological staining (H E) of induced RD and PVR in retina treated with OM-101. (a) Histological staining (H E) demonstrated the following: left (control/Hepes), normal architecture; middle (dispase + PBS), abnormal morphology with inflammatory cells, fibroblasts, and pigment stain RPE cells through the retina denoting PVR; and right (dispase + OM 101), OM 101 treatment maintained the normal structure of the retina. Scale bar 100 μ m or 200 μ m. (b) Number of mice developing RD or PVR. (c) Severity score of the mice retina measured by thickness of the RD, number of inflammatory cells, loss of normal structure, and the appearance of RPE cells inside the retina ( N = 25). Statistics were computed using Student's t -test (two-tailed distribution equal variance). Data is expressed as the mean ± SD.

    Techniques Used: Staining, Mouse Assay, Two Tailed Test

    8) Product Images from "Conditional knockout of N-Myc and STAT Interactor Disrupts Normal Mammary Development and Enhances Metastatic Ability of Mammary Tumors"

    Article Title: Conditional knockout of N-Myc and STAT Interactor Disrupts Normal Mammary Development and Enhances Metastatic Ability of Mammary Tumors

    Journal: Oncogene

    doi: 10.1038/s41388-017-0037-7

    Carcinogen and MMTV-Neu induced Nmi null tumors exhibit invasive morphological features. a Schematic for establishing 3D organoid cultures: tumors from control and knockout models were excised and digested using collagenase (10 mg/mL) and pronase (0.1 mg/mL) at 37 °C to separate tumor cells from the extracellular matrix. Organoids from tumors were overlaid onto 3D matrix (cultrex) in chamber slides (Millipore). b Morphologic behavior of control and Nmi knockout tumor cells was studied in 2 or 3-dimensional growth conditions. For 2D growth, tumor cells were allowed to attach to culture dishes and imaged prior to passaging. The 3D organoid cultures were established as described in the schematic a . Photomicrographs show phase contrast images of 2D and 3D cultures. The 3D culture images were captured after 14 days of culture to examine invasive processes. Scale bar = 100 μM for 2D cultures and 25 μM for 3D cultures. Green arrows in 2D images indicate smooth/cohesive edges of colonies whereas red arrows indicate non cohesive/serrated edges. Branches/organoid in 3D culture were counted and represented as bar graphs * p = 0.002; N = 6 ko mice and 4 cntrl mice (MMTV-Neu) and * p = 0.0001; N = 20 organoids per group (DMBA/MPA). c Organoid cultures from MMTV-Neu Nmi fl/fl and MMTV-Neu Nmi −/− were stained for fibrillar actin (phalloidin) to visualize invasive processes. DAPI was used to stain nuclei. Representative confocal images of organoids from control and knockout models are presented. d Tumor cells from MMTV-Neu control and knockout mice were separated into single cells using 0.25% trypsin/EDTA and dispase II (5 mg/ml) and placed (50,000 cells) into Boyden chambers using fibronectin (10 µg/ml) as a chemoattractant. Cells were allowed 24 h to invade through the matrigel coated chambers, fixed with 4% PFA and stained with crystal violet (the experiment was repeated independently) N = 5 control tumors and 4 knockout tumors. Quantification of invasion is a measure of the percentage of cells that invaded. * p = 0.001
    Figure Legend Snippet: Carcinogen and MMTV-Neu induced Nmi null tumors exhibit invasive morphological features. a Schematic for establishing 3D organoid cultures: tumors from control and knockout models were excised and digested using collagenase (10 mg/mL) and pronase (0.1 mg/mL) at 37 °C to separate tumor cells from the extracellular matrix. Organoids from tumors were overlaid onto 3D matrix (cultrex) in chamber slides (Millipore). b Morphologic behavior of control and Nmi knockout tumor cells was studied in 2 or 3-dimensional growth conditions. For 2D growth, tumor cells were allowed to attach to culture dishes and imaged prior to passaging. The 3D organoid cultures were established as described in the schematic a . Photomicrographs show phase contrast images of 2D and 3D cultures. The 3D culture images were captured after 14 days of culture to examine invasive processes. Scale bar = 100 μM for 2D cultures and 25 μM for 3D cultures. Green arrows in 2D images indicate smooth/cohesive edges of colonies whereas red arrows indicate non cohesive/serrated edges. Branches/organoid in 3D culture were counted and represented as bar graphs * p = 0.002; N = 6 ko mice and 4 cntrl mice (MMTV-Neu) and * p = 0.0001; N = 20 organoids per group (DMBA/MPA). c Organoid cultures from MMTV-Neu Nmi fl/fl and MMTV-Neu Nmi −/− were stained for fibrillar actin (phalloidin) to visualize invasive processes. DAPI was used to stain nuclei. Representative confocal images of organoids from control and knockout models are presented. d Tumor cells from MMTV-Neu control and knockout mice were separated into single cells using 0.25% trypsin/EDTA and dispase II (5 mg/ml) and placed (50,000 cells) into Boyden chambers using fibronectin (10 µg/ml) as a chemoattractant. Cells were allowed 24 h to invade through the matrigel coated chambers, fixed with 4% PFA and stained with crystal violet (the experiment was repeated independently) N = 5 control tumors and 4 knockout tumors. Quantification of invasion is a measure of the percentage of cells that invaded. * p = 0.001

    Techniques Used: Knock-Out, Passaging, Mouse Assay, Staining

    9) Product Images from "Conditional knockout of N-Myc and STAT Interactor Disrupts Normal Mammary Development and Enhances Metastatic Ability of Mammary Tumors"

    Article Title: Conditional knockout of N-Myc and STAT Interactor Disrupts Normal Mammary Development and Enhances Metastatic Ability of Mammary Tumors

    Journal: Oncogene

    doi: 10.1038/s41388-017-0037-7

    Carcinogen and MMTV-Neu induced Nmi null tumors exhibit invasive morphological features. a Schematic for establishing 3D organoid cultures: tumors from control and knockout models were excised and digested using collagenase (10 mg/mL) and pronase (0.1 mg/mL) at 37 °C to separate tumor cells from the extracellular matrix. Organoids from tumors were overlaid onto 3D matrix (cultrex) in chamber slides (Millipore). b Morphologic behavior of control and Nmi knockout tumor cells was studied in 2 or 3-dimensional growth conditions. For 2D growth, tumor cells were allowed to attach to culture dishes and imaged prior to passaging. The 3D organoid cultures were established as described in the schematic a . Photomicrographs show phase contrast images of 2D and 3D cultures. The 3D culture images were captured after 14 days of culture to examine invasive processes. Scale bar = 100 μM for 2D cultures and 25 μM for 3D cultures. Green arrows in 2D images indicate smooth/cohesive edges of colonies whereas red arrows indicate non cohesive/serrated edges. Branches/organoid in 3D culture were counted and represented as bar graphs * p = 0.002; N = 6 ko mice and 4 cntrl mice (MMTV-Neu) and * p = 0.0001; N = 20 organoids per group (DMBA/MPA). c Organoid cultures from MMTV-Neu Nmi fl/fl and MMTV-Neu Nmi −/− were stained for fibrillar actin (phalloidin) to visualize invasive processes. DAPI was used to stain nuclei. Representative confocal images of organoids from control and knockout models are presented. d Tumor cells from MMTV-Neu control and knockout mice were separated into single cells using 0.25% trypsin/EDTA and dispase II (5 mg/ml) and placed (50,000 cells) into Boyden chambers using fibronectin (10 µg/ml) as a chemoattractant. Cells were allowed 24 h to invade through the matrigel coated chambers, fixed with 4% PFA and stained with crystal violet (the experiment was repeated independently) N = 5 control tumors and 4 knockout tumors. Quantification of invasion is a measure of the percentage of cells that invaded. * p = 0.001
    Figure Legend Snippet: Carcinogen and MMTV-Neu induced Nmi null tumors exhibit invasive morphological features. a Schematic for establishing 3D organoid cultures: tumors from control and knockout models were excised and digested using collagenase (10 mg/mL) and pronase (0.1 mg/mL) at 37 °C to separate tumor cells from the extracellular matrix. Organoids from tumors were overlaid onto 3D matrix (cultrex) in chamber slides (Millipore). b Morphologic behavior of control and Nmi knockout tumor cells was studied in 2 or 3-dimensional growth conditions. For 2D growth, tumor cells were allowed to attach to culture dishes and imaged prior to passaging. The 3D organoid cultures were established as described in the schematic a . Photomicrographs show phase contrast images of 2D and 3D cultures. The 3D culture images were captured after 14 days of culture to examine invasive processes. Scale bar = 100 μM for 2D cultures and 25 μM for 3D cultures. Green arrows in 2D images indicate smooth/cohesive edges of colonies whereas red arrows indicate non cohesive/serrated edges. Branches/organoid in 3D culture were counted and represented as bar graphs * p = 0.002; N = 6 ko mice and 4 cntrl mice (MMTV-Neu) and * p = 0.0001; N = 20 organoids per group (DMBA/MPA). c Organoid cultures from MMTV-Neu Nmi fl/fl and MMTV-Neu Nmi −/− were stained for fibrillar actin (phalloidin) to visualize invasive processes. DAPI was used to stain nuclei. Representative confocal images of organoids from control and knockout models are presented. d Tumor cells from MMTV-Neu control and knockout mice were separated into single cells using 0.25% trypsin/EDTA and dispase II (5 mg/ml) and placed (50,000 cells) into Boyden chambers using fibronectin (10 µg/ml) as a chemoattractant. Cells were allowed 24 h to invade through the matrigel coated chambers, fixed with 4% PFA and stained with crystal violet (the experiment was repeated independently) N = 5 control tumors and 4 knockout tumors. Quantification of invasion is a measure of the percentage of cells that invaded. * p = 0.001

    Techniques Used: Knock-Out, Passaging, Mouse Assay, Staining

    10) Product Images from "Hepatocyte Growth Factor Activator Inhibitor Type 1 Maintains the Assembly of Keratin into Desmosomes in Keratinocytes by Regulating Protease-Activated Receptor 2–Dependent p38 Signaling"

    Article Title: Hepatocyte Growth Factor Activator Inhibitor Type 1 Maintains the Assembly of Keratin into Desmosomes in Keratinocytes by Regulating Protease-Activated Receptor 2–Dependent p38 Signaling

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.02.009

    Enhanced vulnerability to mechanical shear stress in hepatocyte growth factor activator inhibitor type 1 (HAI)-1 knockdown (KD) HaCaT. A: Representative images of control (Cont) and HAI-1 KD (KD) HaCaT cells after dispase mechanical dissociation assay. B: Number of fragments generated by mechanical shear stress in the dispase assay. Data are given as means ± SD ( B ). ∗∗ P
    Figure Legend Snippet: Enhanced vulnerability to mechanical shear stress in hepatocyte growth factor activator inhibitor type 1 (HAI)-1 knockdown (KD) HaCaT. A: Representative images of control (Cont) and HAI-1 KD (KD) HaCaT cells after dispase mechanical dissociation assay. B: Number of fragments generated by mechanical shear stress in the dispase assay. Data are given as means ± SD ( B ). ∗∗ P

    Techniques Used: Generated

    11) Product Images from "Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity"

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-25203-3

    Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.
    Figure Legend Snippet: Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Techniques Used: Microscopy

    Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).
    Figure Legend Snippet: Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Techniques Used:

    12) Product Images from "Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus"

    Article Title: Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-03697-7

    Adhesion measurements with modulation of signaling molecules. Dispase-based dissociation assays in HaCaT keratinocytes after applying the specific inhibitors SB202190 (p38MAPK), Bim-X (PKC), U0126 (Erk) or PP2 (Src) for 1 h, followed by either ( a ) 24 h incubation of IgG fractions or ( c ) 2 h incubation of IgG fractions (n = 4; *p ≤ 0.05 vs. c-IgG with the respective mediator). ( b , d ) Representative images of monolayers incubated with 10 μ m MTT to visualize cell fragments.
    Figure Legend Snippet: Adhesion measurements with modulation of signaling molecules. Dispase-based dissociation assays in HaCaT keratinocytes after applying the specific inhibitors SB202190 (p38MAPK), Bim-X (PKC), U0126 (Erk) or PP2 (Src) for 1 h, followed by either ( a ) 24 h incubation of IgG fractions or ( c ) 2 h incubation of IgG fractions (n = 4; *p ≤ 0.05 vs. c-IgG with the respective mediator). ( b , d ) Representative images of monolayers incubated with 10 μ m MTT to visualize cell fragments.

    Techniques Used: Incubation, MTT Assay

    13) Product Images from "Adducin Is Required for Desmosomal Cohesion in Keratinocytes *"

    Article Title: Adducin Is Required for Desmosomal Cohesion in Keratinocytes *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.527127

    Adducin silencing promotes cell dissociation and reduces desmosomal Dsg3 levels. A, after siRNA-mediated silencing of the respective adducin isoform, confluent monolayers were subjected to dispase-based dissociation assays. Specific adducin knockdown
    Figure Legend Snippet: Adducin silencing promotes cell dissociation and reduces desmosomal Dsg3 levels. A, after siRNA-mediated silencing of the respective adducin isoform, confluent monolayers were subjected to dispase-based dissociation assays. Specific adducin knockdown

    Techniques Used:

    The protective effect of Rho-GTPase activation is dependent on both adducin expression and phosphorylation at Ser-726. A, representative Western blot of α- or γ-adducin silencing performed in parallel to dispase-based dissociation assays.
    Figure Legend Snippet: The protective effect of Rho-GTPase activation is dependent on both adducin expression and phosphorylation at Ser-726. A, representative Western blot of α- or γ-adducin silencing performed in parallel to dispase-based dissociation assays.

    Techniques Used: Activation Assay, Expressing, Western Blot

    14) Product Images from "Protective Endogenous Cyclic Adenosine 5?-Monophosphate Signaling Triggered by Pemphigus Autoantibodies"

    Article Title: Protective Endogenous Cyclic Adenosine 5?-Monophosphate Signaling Triggered by Pemphigus Autoantibodies

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1002675

    cAMP increase prevents PV-IgG–mediated loss of cell adhesion and Dsg3 depletion. A, Compared with controls, application of mechanical stress to HaCaT monolayers in dispase-based dissociation assays resulted in pronounced fragment formation following
    Figure Legend Snippet: cAMP increase prevents PV-IgG–mediated loss of cell adhesion and Dsg3 depletion. A, Compared with controls, application of mechanical stress to HaCaT monolayers in dispase-based dissociation assays resulted in pronounced fragment formation following

    Techniques Used:

    15) Product Images from "Dsg2 Upregulation as a Rescue Mechanism in Pemphigus"

    Article Title: Dsg2 Upregulation as a Rescue Mechanism in Pemphigus

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.581370

    Dsg2 is upregulated in murine Dsg3 ko keratinocytes. (A) Dispase-based keratinocyte dissociation assay of stable Dsg3 wt and ko keratinocytes showing compromised intercellular adhesion in cells lacking Dsg3. Columns indicate mean value ± SEM, *P
    Figure Legend Snippet: Dsg2 is upregulated in murine Dsg3 ko keratinocytes. (A) Dispase-based keratinocyte dissociation assay of stable Dsg3 wt and ko keratinocytes showing compromised intercellular adhesion in cells lacking Dsg3. Columns indicate mean value ± SEM, *P

    Techniques Used:

    Heterophilic Dsg2-Dsg3 interactions were less susceptible to autoantibody-induced direct inhibition of Dsg interaction. Binding frequencies of cell-free AFM measurements probing Dsg3-Dsg3 and Dsg2-Dsg3 interaction pairs with and without (ctr) incubation of either a monoclonal anti-Dsg3 antibody (AK23) (A) or PV3-IgG (B) for 1 h (n ≥ 5, each 2500 force-distance curves). Heterophilic Dsg2-Dsg3 interactions are less susceptible against direct inhibition by pathogenic Dsg3 antibodies than homophilic Dsg3 interactions. (C) Dispase-based keratinocyte dissociation assay of Dsg3 wt and ko keratinocytes after treatment with control (ctr) IgG or AK23 for 24 h showing loss of intercellular adhesion after AK23 in wt cells only (n = 3). (D) Keratinocyte dissociation assay of Dsg3 ko keratinocytes after treatment with ctr-IgG, AK23 or an inhibitory anti-Dsg2 antibody (aDsg2) for 24 h revealing drastic effect of aDsg2 antibody in cells lacking Dsg3 (n = 4). Columns indicate mean value normalized to ctr ± SEM, *P
    Figure Legend Snippet: Heterophilic Dsg2-Dsg3 interactions were less susceptible to autoantibody-induced direct inhibition of Dsg interaction. Binding frequencies of cell-free AFM measurements probing Dsg3-Dsg3 and Dsg2-Dsg3 interaction pairs with and without (ctr) incubation of either a monoclonal anti-Dsg3 antibody (AK23) (A) or PV3-IgG (B) for 1 h (n ≥ 5, each 2500 force-distance curves). Heterophilic Dsg2-Dsg3 interactions are less susceptible against direct inhibition by pathogenic Dsg3 antibodies than homophilic Dsg3 interactions. (C) Dispase-based keratinocyte dissociation assay of Dsg3 wt and ko keratinocytes after treatment with control (ctr) IgG or AK23 for 24 h showing loss of intercellular adhesion after AK23 in wt cells only (n = 3). (D) Keratinocyte dissociation assay of Dsg3 ko keratinocytes after treatment with ctr-IgG, AK23 or an inhibitory anti-Dsg2 antibody (aDsg2) for 24 h revealing drastic effect of aDsg2 antibody in cells lacking Dsg3 (n = 4). Columns indicate mean value normalized to ctr ± SEM, *P

    Techniques Used: Inhibition, Binding Assay, Incubation

    16) Product Images from "The Extent of Desmoglein 3 Depletion in Pemphigus Vulgaris Is Dependent on Ca2+-Induced Differentiation"

    Article Title: The Extent of Desmoglein 3 Depletion in Pemphigus Vulgaris Is Dependent on Ca2+-Induced Differentiation

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2011.06.043

    Loss of cell adhesion is blunted by PKC inhibition in HaCaT cells cultured for 3 days and NHEK cells. Dispase-based keratinocyte dissociation assays were performed to quantify loss of cell adhesion. Treatment with PV1-IgG for 24 hours induced strong fragmentation under all conditions. Simultaneous treatment with Gö6976 or safingol blunted cell dissociation in HaCaT cells cultured for 3 days ( A ) and in NHEK cells ( C ) but was ineffective in HaCaT cells cultured for 8 days ( B ) ( n = > 6). * P
    Figure Legend Snippet: Loss of cell adhesion is blunted by PKC inhibition in HaCaT cells cultured for 3 days and NHEK cells. Dispase-based keratinocyte dissociation assays were performed to quantify loss of cell adhesion. Treatment with PV1-IgG for 24 hours induced strong fragmentation under all conditions. Simultaneous treatment with Gö6976 or safingol blunted cell dissociation in HaCaT cells cultured for 3 days ( A ) and in NHEK cells ( C ) but was ineffective in HaCaT cells cultured for 8 days ( B ) ( n = > 6). * P

    Techniques Used: Inhibition, Cell Culture

    17) Product Images from "RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells"

    Article Title: RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-020-01976-1

    Cell morphology and adhesion alterations. a Immunofluorescence of OCT4 expression and localization in hHFMSCs, adherent hHFMSCs OCT4 , and floating hHFMSCs OCT4 . Adherent, adherent hHFMSCs OCT4 ; Floating, floating hHFMSCs OCT4 ; NC, negative control. Red represents OCT4, blue for DAPI and green for EGFP. b The morphology of hHFMSCs was gradually altered after the transduction of OCT4. Yellow arrows indicate round floating hHFMSCs OCT4 . The first four upper pictures were taken to observe the morphology of the cells under an optical microscope, and the last lower four pictures were taken to observe the expression of EGFP after OCT4 transduction under a fluorescence microscope. These pictures were all taken from randomly selected fields. c Flow cytometric plot of three groups of hHFMSCs and geometric mean of the FSC. d Percentage of single cells adhered to the plates after treatment with dispase. The percentage of disassociated individual cells to the total number of cells is negatively correlated with cell-to-cell adhesion. e Percentage of cells adhered to the Matrigel-coated plates after rinsing with PBS three times. * P
    Figure Legend Snippet: Cell morphology and adhesion alterations. a Immunofluorescence of OCT4 expression and localization in hHFMSCs, adherent hHFMSCs OCT4 , and floating hHFMSCs OCT4 . Adherent, adherent hHFMSCs OCT4 ; Floating, floating hHFMSCs OCT4 ; NC, negative control. Red represents OCT4, blue for DAPI and green for EGFP. b The morphology of hHFMSCs was gradually altered after the transduction of OCT4. Yellow arrows indicate round floating hHFMSCs OCT4 . The first four upper pictures were taken to observe the morphology of the cells under an optical microscope, and the last lower four pictures were taken to observe the expression of EGFP after OCT4 transduction under a fluorescence microscope. These pictures were all taken from randomly selected fields. c Flow cytometric plot of three groups of hHFMSCs and geometric mean of the FSC. d Percentage of single cells adhered to the plates after treatment with dispase. The percentage of disassociated individual cells to the total number of cells is negatively correlated with cell-to-cell adhesion. e Percentage of cells adhered to the Matrigel-coated plates after rinsing with PBS three times. * P

    Techniques Used: Immunofluorescence, Expressing, Negative Control, Transduction, Microscopy, Fluorescence

    18) Product Images from "A non-aggressive, highly efficient, enzymatic method for dissociation of human brain-tumors and brain-tissues to viable single-cells"

    Article Title: A non-aggressive, highly efficient, enzymatic method for dissociation of human brain-tumors and brain-tissues to viable single-cells

    Journal: BMC Neuroscience

    doi: 10.1186/s12868-016-0262-y

    BT dissociation to single cells using dispase (Disp) or neutral protease (NP). a Cellular viability and b dissociation quality (CG) of BTs dissociated with dispase or NP, at the respective enzyme’s optimal dissociation time. Following indicated times (1 or 2 h) the cells were triturated using a Pasteur pipette and their viability and CG were determined. Oli—Oligodenderoglioma, OliAst–Oligoastrocytoma, Ast–Astrocytoma, GBM–Glioblastoma, Mets–Metastasis to the brain (lung–lun and melanoma–Mel), Epi + Br—Epileptic foci, and peritumoral brain tissue. Parenthesis indicate the grade of the tumors, e.g. Oli(2–3). Statistics Viability following dissociation of all glial tumors (Oli, OliAst, Ast, GBM) using NP-2 h to dispase-1 h (P
    Figure Legend Snippet: BT dissociation to single cells using dispase (Disp) or neutral protease (NP). a Cellular viability and b dissociation quality (CG) of BTs dissociated with dispase or NP, at the respective enzyme’s optimal dissociation time. Following indicated times (1 or 2 h) the cells were triturated using a Pasteur pipette and their viability and CG were determined. Oli—Oligodenderoglioma, OliAst–Oligoastrocytoma, Ast–Astrocytoma, GBM–Glioblastoma, Mets–Metastasis to the brain (lung–lun and melanoma–Mel), Epi + Br—Epileptic foci, and peritumoral brain tissue. Parenthesis indicate the grade of the tumors, e.g. Oli(2–3). Statistics Viability following dissociation of all glial tumors (Oli, OliAst, Ast, GBM) using NP-2 h to dispase-1 h (P

    Techniques Used: Transferring, AST Assay

    BT dissociation ON to single cells using dispase or NP. a Cellular viability and b dissociation quality of NP-2 h versus NP-ON. c Cellular viability and d dissociation quality of dispase-1 h versus dispase-ON. BTs were dissociated for 1–2 h or ON. Following indicated times, the cells were triturated and their viability and CG was determined. Statistics Viability of dissociated cells and the dissociation quality was not different for all glial tumors between NP 2 h to NP-ON, or between dispase 1 h to dispase-ON
    Figure Legend Snippet: BT dissociation ON to single cells using dispase or NP. a Cellular viability and b dissociation quality of NP-2 h versus NP-ON. c Cellular viability and d dissociation quality of dispase-1 h versus dispase-ON. BTs were dissociated for 1–2 h or ON. Following indicated times, the cells were triturated and their viability and CG was determined. Statistics Viability of dissociated cells and the dissociation quality was not different for all glial tumors between NP 2 h to NP-ON, or between dispase 1 h to dispase-ON

    Techniques Used:

    Brain tumor (BT) dissociation to single cells using various enzymes. a Cellular viability and b dissociation cumulative grade (CG) for BTs dissociated with dispase (Disp), papain, a combination of DNase, collagenase and hyaluronidase (DCH), or mechanical dissociation only (none). See text for calculation of CG. Primary brain tumors were dissociated to single cells for 1 hour (1 h), 2 hour (2 h), or overnight (ON) at optimal enzyme concentrations—(see text). After the indicated times, the cells were triturated using a Pasteur pipette and their viability and CG was determined. Statistics: Viability of Disp or DCH dissociated-tumors to mechanically dissociated tumors (P
    Figure Legend Snippet: Brain tumor (BT) dissociation to single cells using various enzymes. a Cellular viability and b dissociation cumulative grade (CG) for BTs dissociated with dispase (Disp), papain, a combination of DNase, collagenase and hyaluronidase (DCH), or mechanical dissociation only (none). See text for calculation of CG. Primary brain tumors were dissociated to single cells for 1 hour (1 h), 2 hour (2 h), or overnight (ON) at optimal enzyme concentrations—(see text). After the indicated times, the cells were triturated using a Pasteur pipette and their viability and CG was determined. Statistics: Viability of Disp or DCH dissociated-tumors to mechanically dissociated tumors (P

    Techniques Used: Transferring

    19) Product Images from "SOXF factors regulate murine satellite cell self-renewal and function through inhibition of β-catenin activity"

    Article Title: SOXF factors regulate murine satellite cell self-renewal and function through inhibition of β-catenin activity

    Journal: eLife

    doi: 10.7554/eLife.26039

    Minimal CD31+ cell contamination in FACS-isolated skeletal muscle stem cells. Pax3 GFP/+ trunk muscles from adult mice were digested in a solution of collagenase/dispase, filtered, and immunolabeled for the endothelial cell marker CD31-PE (Phycoerythrin fluorochrome) before FACS. ( A ) Gating for CD31-PE/GFP. ( B ) Gating for single cell (SSC-side scatter)/GFP. ( C ) Histograms and gating for cell number/CD31+ cells in GFP- and GFP+ cell fractions. ( D ) Graphic illustrating the proportion of CD31+ cells. n = 3. Data expressed as mean ± s.e.m.
    Figure Legend Snippet: Minimal CD31+ cell contamination in FACS-isolated skeletal muscle stem cells. Pax3 GFP/+ trunk muscles from adult mice were digested in a solution of collagenase/dispase, filtered, and immunolabeled for the endothelial cell marker CD31-PE (Phycoerythrin fluorochrome) before FACS. ( A ) Gating for CD31-PE/GFP. ( B ) Gating for single cell (SSC-side scatter)/GFP. ( C ) Histograms and gating for cell number/CD31+ cells in GFP- and GFP+ cell fractions. ( D ) Graphic illustrating the proportion of CD31+ cells. n = 3. Data expressed as mean ± s.e.m.

    Techniques Used: FACS, Isolation, Mouse Assay, Immunolabeling, Marker

    20) Product Images from "Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity"

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-25203-3

    Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.
    Figure Legend Snippet: Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Techniques Used: Microscopy

    Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).
    Figure Legend Snippet: Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Techniques Used:

    Related Articles

    Incubation:

    Article Title: Dsg2 Upregulation as a Rescue Mechanism in Pemphigus
    Article Snippet: .. Briefly, epidermis was obtained from neonatal mice by incubation of the skin with Dispase II (Sigma-Aldrich, Munich, Germany) overnight. .. Epidermal cells were isolated after treatment for 1 h with Accutase (Sigma-Aldrich, Munich, Germany) and seeded in complete FAD media (0.05mM CaCl2 , PAN Biotech, Aidenbach, Germany) on collagen I (rat tail; BD Bioscience, New Jersey, US) coated flasks and maintained at 35°C and 5% CO2 .

    Article Title: Desmocollin-2 affects the adhesive strength and cytoskeletal arrangement in esophageal squamous cell carcinoma cells
    Article Snippet: .. Subsequently, 24 h after reaching confluency, the cultures were washed twice in phosphate-buffered saline (PBS) and incubated in 1 ml dispase (2.4 U/ml; Sigma) for 30 min, as previously described ( ). ..

    Article Title: Pemphigus Vulgaris IgG Cause Loss of Desmoglein-Mediated Adhesion and Keratinocyte Dissociation Independent of Epidermal Growth Factor Receptor
    Article Snippet: .. After incubation for 24 hours under various conditions, cells were washed with Hanks’ buffered salt solution and treated for 30 minutes with 0.3 ml dispase II (2.4 U/ml, Sigma) at 37°C. ..

    Article Title: Arrhythmogenic cardiomyopathy related DSG2 mutations affect desmosomal cadherin binding kinetics
    Article Snippet: .. After two washes with phosphate-buffered saline, the adherent cells were incubated at 37 °C for 20 minutes with 0.3 ml dispase II (2.4 U/ml, Sigma Aldrich, Darmstadt, Germany) resulting in a non-adherent cell monolayer. ..

    Injection:

    Article Title: OM-101 Decreases the Fibrotic Response Associated with Proliferative Vitreoretinopathy
    Article Snippet: .. Then, 3 μ l of Dispase II (10 mg/ml, Sigma number D493, dissolved in Hepes/KOH pH = 7.4, 50 mM NaCl buffer) was injected intravitreally to the right eye. .. As control, mice in group A were injected with 3 μ l of the Hepes solution (N = 5 mice).

    Mouse Assay:

    Article Title: Dsg2 Upregulation as a Rescue Mechanism in Pemphigus
    Article Snippet: .. Briefly, epidermis was obtained from neonatal mice by incubation of the skin with Dispase II (Sigma-Aldrich, Munich, Germany) overnight. .. Epidermal cells were isolated after treatment for 1 h with Accutase (Sigma-Aldrich, Munich, Germany) and seeded in complete FAD media (0.05mM CaCl2 , PAN Biotech, Aidenbach, Germany) on collagen I (rat tail; BD Bioscience, New Jersey, US) coated flasks and maintained at 35°C and 5% CO2 .

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  • 99
    Millipore dispase
    hES cells cultured on fibronectin in hESF9 medium. Phase-contrast microphotographs of KhES-1 at passage 16 and KhES-3 at passage 6 cultured on fibronectin in hESF9 medium. After 2 d, the KhES-1 cells and KhES-3 cells (from Kyoto University) cultured on MEF in KSR-based medium were passaged by the routine procedure onto the new MEF; the medium was changed to hESF9 medium, and then the cells were cultured on MEF for more than 4 d. Previously, we dissociated the hES cells with EDTA, but some of cell lines could not survive after the dissociation with EDTA. Then, we have utilized 1 U/ml <t>dispase</t> (Roche Applied Science) for approximately 1 min at 37°C to dissociate the hES/iPS colonies into the small clumps. The cells were washed twice at 20 g and then seeded on fibronectin-coated flask (Corning Costar) in hESF9 medium.
    Dispase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 391 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore dispase ii
    Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by <t>dispase</t> and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.
    Dispase Ii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dispase ii/product/Millipore
    Average 99 stars, based on 196 article reviews
    Price from $9.99 to $1999.99
    dispase ii - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier


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    hES cells cultured on fibronectin in hESF9 medium. Phase-contrast microphotographs of KhES-1 at passage 16 and KhES-3 at passage 6 cultured on fibronectin in hESF9 medium. After 2 d, the KhES-1 cells and KhES-3 cells (from Kyoto University) cultured on MEF in KSR-based medium were passaged by the routine procedure onto the new MEF; the medium was changed to hESF9 medium, and then the cells were cultured on MEF for more than 4 d. Previously, we dissociated the hES cells with EDTA, but some of cell lines could not survive after the dissociation with EDTA. Then, we have utilized 1 U/ml dispase (Roche Applied Science) for approximately 1 min at 37°C to dissociate the hES/iPS colonies into the small clumps. The cells were washed twice at 20 g and then seeded on fibronectin-coated flask (Corning Costar) in hESF9 medium.

    Journal: In Vitro Cellular & Developmental Biology. Animal

    Article Title: Advantages and difficulties in culturing human pluripotent stem cells in growth factor-defined serum-free medium

    doi: 10.1007/s11626-010-9317-z

    Figure Lengend Snippet: hES cells cultured on fibronectin in hESF9 medium. Phase-contrast microphotographs of KhES-1 at passage 16 and KhES-3 at passage 6 cultured on fibronectin in hESF9 medium. After 2 d, the KhES-1 cells and KhES-3 cells (from Kyoto University) cultured on MEF in KSR-based medium were passaged by the routine procedure onto the new MEF; the medium was changed to hESF9 medium, and then the cells were cultured on MEF for more than 4 d. Previously, we dissociated the hES cells with EDTA, but some of cell lines could not survive after the dissociation with EDTA. Then, we have utilized 1 U/ml dispase (Roche Applied Science) for approximately 1 min at 37°C to dissociate the hES/iPS colonies into the small clumps. The cells were washed twice at 20 g and then seeded on fibronectin-coated flask (Corning Costar) in hESF9 medium.

    Article Snippet: We have used 1 U/ml dispase (Roche Applied Science, Indianapolis, IN) to dissociate the cell colonies and washed the dispase with the medium supplemented with recombinant human albumin (1 mg/ml, Millipore, Bedford, MA).

    Techniques: Cell Culture

    Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Journal: Scientific Reports

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    doi: 10.1038/s41598-018-25203-3

    Figure Lengend Snippet: Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Article Snippet: Dispase II was purchased from EMD Millipore.

    Techniques: Microscopy

    Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Journal: Scientific Reports

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    doi: 10.1038/s41598-018-25203-3

    Figure Lengend Snippet: Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Article Snippet: Dispase II was purchased from EMD Millipore.

    Techniques:

    Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Journal: Scientific Reports

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    doi: 10.1038/s41598-018-25203-3

    Figure Lengend Snippet: Contrast microscope images of HCT-116 tumor spheroids formation process. ( A ) HCT-116 cell sheet (Sheet growth = 10 days) on petri dish and ( B – E ) cell sheet was detached by dispase and kept orbital-shaking at 30 rph with dispase doped medium (1/6) for different time. All scale bars are 200 µm.

    Article Snippet: Dispase II was purchased from EMD Millipore.

    Techniques: Microscopy

    Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Journal: Scientific Reports

    Article Title: Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

    doi: 10.1038/s41598-018-25203-3

    Figure Lengend Snippet: Cartoon illustration of cellular spheroid formation. Single cells are seeded onto petri dishes ( A ), and after a few days of growth, cell sheets form ( B ). Cell sheets can be detached from the petri dishes by dispase and the sheets can be maintained ( C ). Shaking the cell sheets in dispase-doped media allows their edges fold inward ( D ) and eventually the cell sheets become spheroids ( E ).

    Article Snippet: Dispase II was purchased from EMD Millipore.

    Techniques: