Structured Review

Roche collagenase
Collagenase, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 452 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/collagenase/product/Roche
Average 99 stars, based on 452 article reviews
Price from $9.99 to $1999.99
collagenase - by Bioz Stars, 2020-05
99/100 stars

Images

Related Articles

Flow Cytometry:

Article Title: Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis
Article Snippet: .. Preparation of the Mammary Epithelial Cells and Flow Cytometry Mammary fat pads were mechanically dissociated with scissors and digested for 90 min at 37°C in CO2 -independent medium (Invitrogen) supplemented with 5% fetal bovine serum, 3 mg/ml collagenase (Roche Diagnostics) and 100 U/ml hyaluronidase (Sigma). .. The resultant suspension was sequentially resuspended in 0.25% trypsin–EDTA for 1 min, and then 5 min in 5 mg/ml dispase (Roche Diagnostics) with 0.1 mg ml–1 DNase I (Sigma) followed by filtration trough a 40 µm mesh.

Article Title: Acute Lung Injury and Acute Kidney Injury Are Established by Four Hours in Experimental Sepsis and Are Improved with Pre, but Not Post, Sepsis Administration of TNF-? Antibodies
Article Snippet: .. Flow cytometry for lung neutrophils Lung parenchyma was minced and processed by enzymatic digestion: 2 mg/ml collagenase (Roche) in RPMI media (Gibco), incubated, strained, and RBCs were lysed with ACK red blood cell lysis buffer (Quality Biological), and washed. .. Cells were stained with: CD11c-PECy7, CD11b-Pacific blue, F4/80-PerCP-Cy5 (eBiosciences), CD45-V500, and Ly6G-APCCy7 (BD Biosciences) and fixed in 200 µl of 1% paraformaldehyde (Sigma).

Isolation:

Article Title: Altered Pancreatic Islet Function and Morphology in Mice Lacking the Beta-Cell Surface Protein Neuroligin-2
Article Snippet: .. Islets were isolated from WT and neuroligin-2 knockout (KO) mice and also from Sprague-Dawley rats (240–260 g; Charles River Laboratories) using collagenase (Liberase, Roche) as previously described , , . ..

Cytometry:

Article Title: Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis
Article Snippet: .. Preparation of the Mammary Epithelial Cells and Flow Cytometry Mammary fat pads were mechanically dissociated with scissors and digested for 90 min at 37°C in CO2 -independent medium (Invitrogen) supplemented with 5% fetal bovine serum, 3 mg/ml collagenase (Roche Diagnostics) and 100 U/ml hyaluronidase (Sigma). .. The resultant suspension was sequentially resuspended in 0.25% trypsin–EDTA for 1 min, and then 5 min in 5 mg/ml dispase (Roche Diagnostics) with 0.1 mg ml–1 DNase I (Sigma) followed by filtration trough a 40 µm mesh.

Article Title: Acute Lung Injury and Acute Kidney Injury Are Established by Four Hours in Experimental Sepsis and Are Improved with Pre, but Not Post, Sepsis Administration of TNF-? Antibodies
Article Snippet: .. Flow cytometry for lung neutrophils Lung parenchyma was minced and processed by enzymatic digestion: 2 mg/ml collagenase (Roche) in RPMI media (Gibco), incubated, strained, and RBCs were lysed with ACK red blood cell lysis buffer (Quality Biological), and washed. .. Cells were stained with: CD11c-PECy7, CD11b-Pacific blue, F4/80-PerCP-Cy5 (eBiosciences), CD45-V500, and Ly6G-APCCy7 (BD Biosciences) and fixed in 200 µl of 1% paraformaldehyde (Sigma).

Knock-Out:

Article Title: Altered Pancreatic Islet Function and Morphology in Mice Lacking the Beta-Cell Surface Protein Neuroligin-2
Article Snippet: .. Islets were isolated from WT and neuroligin-2 knockout (KO) mice and also from Sprague-Dawley rats (240–260 g; Charles River Laboratories) using collagenase (Liberase, Roche) as previously described , , . ..

Mouse Assay:

Article Title: Altered Pancreatic Islet Function and Morphology in Mice Lacking the Beta-Cell Surface Protein Neuroligin-2
Article Snippet: .. Islets were isolated from WT and neuroligin-2 knockout (KO) mice and also from Sprague-Dawley rats (240–260 g; Charles River Laboratories) using collagenase (Liberase, Roche) as previously described , , . ..

Article Title: Emerging Role of HMGB1 in the Pathogenesis of Schistosomiasis Liver Fibrosis
Article Snippet: .. Mice were perfused through the portal vein with HANKS A and then HANKS B medium containing 0.05% collagenase (Roche Applied Science, Indianapolis, IN) and the perfused liver tissue was passed through a 40-μm nylon mesh filter ( , ). .. Primary hepatocytes were cultured at 37°C in 5% CO2 /95% O2 in Williams' medium E (Sigma-Aldrich, Missouri, USA) containing 10% fetal bovine serum (Gibco), 50 units/mL penicillin, 50 g/mL streptomycin (Sigma-Aldrich, Missouri, USA) were plated on collagen-coated coverslips (50 μg/mL) (BD Biosciences, San Jose, CA).

Article Title: Prenatal Betamethasone interferes with immune system development and alters target cells in autoimmune diabetes
Article Snippet: .. To isolate islets, pancreases from NOD mice were perfused with collagenase (Collagenase P, Roche, Indianapolis, IN) through the common bile duct . ..

Incubation:

Article Title: Chronic inflammatory injury results in increased coupling of delta opioid receptors to voltage-gated Ca2+ channels
Article Snippet: .. The DRGs were collected in Complete Saline Solution (CSS; in mM, NaCl: 137, KCl: 5.3, MgCl2 :1, Sorbitol: 25, HEPES: 10, CaCl2 : 3) and incubated in collagenase (1.25U of TH, Roche, Indianopolis, IN), 250 nm EDTA for 20 min at 32 C, transferred to fresh CSS containing collagenase (1.25U of TM, Roche) with 250 nm EDTA and 0.25U papain (Roche) and incubated for 10 min at 32 C. After 2 washes and physical trituration through a series of graded Pasteur pipettes, the cells were spun (1000 rpm, 3 min) and plated in Neurobasal /B27/Glumax/Antibiotic/Antimycotic (Life Technologies, Grand Island, NY) supplemented with 10 ng/ml NGF (Life Technologies). ..

Article Title: Acute Lung Injury and Acute Kidney Injury Are Established by Four Hours in Experimental Sepsis and Are Improved with Pre, but Not Post, Sepsis Administration of TNF-? Antibodies
Article Snippet: .. Flow cytometry for lung neutrophils Lung parenchyma was minced and processed by enzymatic digestion: 2 mg/ml collagenase (Roche) in RPMI media (Gibco), incubated, strained, and RBCs were lysed with ACK red blood cell lysis buffer (Quality Biological), and washed. .. Cells were stained with: CD11c-PECy7, CD11b-Pacific blue, F4/80-PerCP-Cy5 (eBiosciences), CD45-V500, and Ly6G-APCCy7 (BD Biosciences) and fixed in 200 µl of 1% paraformaldehyde (Sigma).

Lysis:

Article Title: Acute Lung Injury and Acute Kidney Injury Are Established by Four Hours in Experimental Sepsis and Are Improved with Pre, but Not Post, Sepsis Administration of TNF-? Antibodies
Article Snippet: .. Flow cytometry for lung neutrophils Lung parenchyma was minced and processed by enzymatic digestion: 2 mg/ml collagenase (Roche) in RPMI media (Gibco), incubated, strained, and RBCs were lysed with ACK red blood cell lysis buffer (Quality Biological), and washed. .. Cells were stained with: CD11c-PECy7, CD11b-Pacific blue, F4/80-PerCP-Cy5 (eBiosciences), CD45-V500, and Ly6G-APCCy7 (BD Biosciences) and fixed in 200 µl of 1% paraformaldehyde (Sigma).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89
    Roche collagenase d solution
    Increased B-cell infiltration and TGF-β expression on TIL-B in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, day 15 and day 21, spleen, TDLN and tumor tissue were processed and stained for the indicated markers on B cells. (A) Kinetics of B-cell infiltration into tumor tissues. At the indicated days following implantation, excised tumors were digested with <t>Collagenase</t> D and single-cell suspensions prepared as outlined in Methods. CD45 + infiltrating lymphocytes were analyzed for CD19 expression (EMT-6 tumor cells do not express CD45). Flow data represent one of three experiments and 3~5 mice per group, mean ± SD. (B) TIL-B phenotype in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, and day 21, spleen, TDLN and tumor tissues were processed and stained for the indicated markers on B cells. CD45 + CD19 + cells were analyzed for surface marker expression as shown. Representative sampling of one mouse at each time point is shown. (C) LAP/TGF-β expression on day 21 on CD19 + splenic B cells, TDLN CD19 + cells and CD19 + TIL-B cells. Twenty-one days post tumor implantation. 5 mice per group, mean ± SD. Statistical analysis using paired t -test, P ≤ 0.0002 for tumor-infiltrating LAP/TGF-β + B cells compared to LAP/TGF-β + splenic or TDLN B cells. (D) LAP/TGF-β expression on day 8, and day 21 on CD19 + TIL-B cells. 5 mice per group, mean ± SD. (E) TGF-β and IL-10 co-expression on splenic and TIL-B: 30 days post EMT-6 implantation, spleen and tumor tissue were processed as described in Methods. Isolated cells were stimulated with LPS overnight and re-stimulated with PMA/ION+ LPS+ Brefeldin A for 6h and stained for intracellular IL-10 and membrane LAP/TGF-β expression. The frequency of LAP/TGF-β1 + CD19 + cells within the CD45 + population is indicated. 5 mice per group, mean ± SD. Flow data from a representative mouse are shown.
    Collagenase D Solution, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase d solution/product/Roche
    Average 89 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    collagenase d solution - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    92
    Roche collagenase gold plus bp protease vitacyte
    Islet yields for the three enzymes used. <t>VitaCyte</t> (n = 8), Roche (n = 48), and Serva (n = 15). Total IEQ: pre-purification (A); post-purification (B); post-culture (C). Results for IEQ/g digested pancreas tissue: pre-purification (D); post-purification (E); post-culture (F). No significant differences were found between enzymes in terms of islet yields expressed in total IEQ or IEQ/g, at pre-purification, post-purification, and post-culture.
    Collagenase Gold Plus Bp Protease Vitacyte, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase gold plus bp protease vitacyte/product/Roche
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    collagenase gold plus bp protease vitacyte - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    90
    Roche collagenase dispase i
    CD83’s TM domain is necessary and sufficient for CD4 T cell selection. (A) Experimental strategy to reconstitute expression of CD83 or chimeric variants thereof in TECs in vivo. WT or Cd83 −/− embryonic TECs, obtained by <t>collagenase/dispase</t> (C/D) digestion and CD45 depletion of E14–16 thymuses, were transduced with vectors encoding equimolar amounts of the construct of interest and EGFP via a T2A linker. RTOCs were prepared by 48-h cultivation on a nylon membrane and subsequently transplanted under the kidney capsule. Thymocyte subsets and TECs were analyzed 5–6 wk after transplantation. sinUTR, self-inactivating UTR; Ubi, ubiquitin; UTR, untranslated region. (B) Expression of the virally encoded GFP reporter in gated cTECs (CD45 − EpCAM + Ly51 + CD80 − ) and mTECs (CD45 − EpCAM + Ly51 − CD80 + ) from RTOCs prepared with a full-length CD83-encoding vector (#2 in F) on day 7 after transplantation. Gray histograms are noninfected control cTECs or mTECs. (C) GFP expression in TECs from RTOCs on days 5 and 35 after transplantation. Gray histograms are TECs from nontransduced RTOCs. (D) CD4 SP thymocyte frequencies in mixed RTOCs with titrated ratios of WT and Cd83 −/− TECs. Individual data points (one or two per time point) are indicated. (E) Thymocyte subsets in Cd83 −/− RTOCs or Cd83 −/− RTOCs from full-length CD83-transduced material. (F) Frequency of CD4 SP cells in RTOCs from Cd83 −/− TECs transduced with viral constructs (Constr.) #1–6 (schematically depicted below the respective data points). The mean transduction efficacy (percentage of GFP + cells among gated CD45 − EpCAM + TECs) ± SEM is shown. n ≥ 3. (G) TECs from RTOCs transduced with the indicated lentiviral constructs as indicated in F were stained for the EC domain of CD83. Unshaded histograms are gated on GFP + TECs (CD45 − EpCAM + cells). Gray histograms are gated on GFP − TECs. The MFI of gated GFP + TECs is indicated. Data in B, C, and E are representative of n ≥ 6 each in at least two independent experiments. In D and F, each data point corresponds to an individual RTOC. ***, P
    Collagenase Dispase I, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase dispase i/product/Roche
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase dispase i - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    95
    Roche collagenase d
    Enriched pancreatic ducts and pancreatic duct epithelium in culture. A: Enriched pancreatic ducts after partial digestion with <t>collagenase</t> D and filtration through 85-μm nylon mesh. Phase contrast, original magnification, ×100. B: Primary culture of duct cells embedded in rat-tail collagen gel with a nylon mesh. Expanding duct epithelium can be seen in the lattice. Original magnification, ×100. C: Electron microscopy of the cultured pancreatic duct cells. Their epithelial nature was apparent by the prominent microvillus processes along portions of the plasma membrane ( thick white arrows ) and formed tight junctions ( solid black arrows ) between the cohered cells. No secretary granules are seen to suggest acinar cell derivation.
    Collagenase D, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 199 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase d/product/Roche
    Average 95 stars, based on 199 article reviews
    Price from $9.99 to $1999.99
    collagenase d - by Bioz Stars, 2020-05
    95/100 stars
      Buy from Supplier

    Image Search Results


    Increased B-cell infiltration and TGF-β expression on TIL-B in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, day 15 and day 21, spleen, TDLN and tumor tissue were processed and stained for the indicated markers on B cells. (A) Kinetics of B-cell infiltration into tumor tissues. At the indicated days following implantation, excised tumors were digested with Collagenase D and single-cell suspensions prepared as outlined in Methods. CD45 + infiltrating lymphocytes were analyzed for CD19 expression (EMT-6 tumor cells do not express CD45). Flow data represent one of three experiments and 3~5 mice per group, mean ± SD. (B) TIL-B phenotype in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, and day 21, spleen, TDLN and tumor tissues were processed and stained for the indicated markers on B cells. CD45 + CD19 + cells were analyzed for surface marker expression as shown. Representative sampling of one mouse at each time point is shown. (C) LAP/TGF-β expression on day 21 on CD19 + splenic B cells, TDLN CD19 + cells and CD19 + TIL-B cells. Twenty-one days post tumor implantation. 5 mice per group, mean ± SD. Statistical analysis using paired t -test, P ≤ 0.0002 for tumor-infiltrating LAP/TGF-β + B cells compared to LAP/TGF-β + splenic or TDLN B cells. (D) LAP/TGF-β expression on day 8, and day 21 on CD19 + TIL-B cells. 5 mice per group, mean ± SD. (E) TGF-β and IL-10 co-expression on splenic and TIL-B: 30 days post EMT-6 implantation, spleen and tumor tissue were processed as described in Methods. Isolated cells were stimulated with LPS overnight and re-stimulated with PMA/ION+ LPS+ Brefeldin A for 6h and stained for intracellular IL-10 and membrane LAP/TGF-β expression. The frequency of LAP/TGF-β1 + CD19 + cells within the CD45 + population is indicated. 5 mice per group, mean ± SD. Flow data from a representative mouse are shown.

    Journal: International Immunology

    Article Title: Mammary-tumor-educated B cells acquire LAP/TGF-β and PD-L1 expression and suppress anti-tumor immune responses

    doi: 10.1093/intimm/dxw007

    Figure Lengend Snippet: Increased B-cell infiltration and TGF-β expression on TIL-B in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, day 15 and day 21, spleen, TDLN and tumor tissue were processed and stained for the indicated markers on B cells. (A) Kinetics of B-cell infiltration into tumor tissues. At the indicated days following implantation, excised tumors were digested with Collagenase D and single-cell suspensions prepared as outlined in Methods. CD45 + infiltrating lymphocytes were analyzed for CD19 expression (EMT-6 tumor cells do not express CD45). Flow data represent one of three experiments and 3~5 mice per group, mean ± SD. (B) TIL-B phenotype in vivo . BALB/c mice were subcutaneously injected with 2×10 5 EMT-6 tumor cells on day 0. At day 8, and day 21, spleen, TDLN and tumor tissues were processed and stained for the indicated markers on B cells. CD45 + CD19 + cells were analyzed for surface marker expression as shown. Representative sampling of one mouse at each time point is shown. (C) LAP/TGF-β expression on day 21 on CD19 + splenic B cells, TDLN CD19 + cells and CD19 + TIL-B cells. Twenty-one days post tumor implantation. 5 mice per group, mean ± SD. Statistical analysis using paired t -test, P ≤ 0.0002 for tumor-infiltrating LAP/TGF-β + B cells compared to LAP/TGF-β + splenic or TDLN B cells. (D) LAP/TGF-β expression on day 8, and day 21 on CD19 + TIL-B cells. 5 mice per group, mean ± SD. (E) TGF-β and IL-10 co-expression on splenic and TIL-B: 30 days post EMT-6 implantation, spleen and tumor tissue were processed as described in Methods. Isolated cells were stimulated with LPS overnight and re-stimulated with PMA/ION+ LPS+ Brefeldin A for 6h and stained for intracellular IL-10 and membrane LAP/TGF-β expression. The frequency of LAP/TGF-β1 + CD19 + cells within the CD45 + population is indicated. 5 mice per group, mean ± SD. Flow data from a representative mouse are shown.

    Article Snippet: Twenty-one days post tumor implantation, tumor tissues were pooled and cut into small pieces and incubated in collagenase D solution (RPMI-1640 supplemented with 2% FBS and 1mg ml–1 of collagenase D, Roche Applied Science) for 30min in a 37°C incubator with gentle shaking.

    Techniques: Expressing, In Vivo, Mouse Assay, Injection, Staining, Flow Cytometry, Marker, Sampling, Tumor Implantation, Isolation

    Islet yields for the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). Total IEQ: pre-purification (A); post-purification (B); post-culture (C). Results for IEQ/g digested pancreas tissue: pre-purification (D); post-purification (E); post-culture (F). No significant differences were found between enzymes in terms of islet yields expressed in total IEQ or IEQ/g, at pre-purification, post-purification, and post-culture.

    Journal: Islets

    Article Title: Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

    doi: 10.1080/19382014.2017.1417716

    Figure Lengend Snippet: Islet yields for the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). Total IEQ: pre-purification (A); post-purification (B); post-culture (C). Results for IEQ/g digested pancreas tissue: pre-purification (D); post-purification (E); post-culture (F). No significant differences were found between enzymes in terms of islet yields expressed in total IEQ or IEQ/g, at pre-purification, post-purification, and post-culture.

    Article Snippet: The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15).

    Techniques: Purification

    Pancreas digestion time (A), digested tissue (B), and digestion percentage (C) for three enzyme groups. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There was no significant difference in digestion time among three groups ( Fig. 1A ). Digested tissue volume showed no significant difference between VitaCyte and Roche, but was significant between VitaCyte and Serva (*p = 0.020)( Fig. 1B ). Digestion percentage showed significant difference between VitaCyte and Roche (*p = 0.026), as well as between VitaCyte and Serva (**p = 0.004) ( Fig. 1C ).

    Journal: Islets

    Article Title: Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

    doi: 10.1080/19382014.2017.1417716

    Figure Lengend Snippet: Pancreas digestion time (A), digested tissue (B), and digestion percentage (C) for three enzyme groups. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There was no significant difference in digestion time among three groups ( Fig. 1A ). Digested tissue volume showed no significant difference between VitaCyte and Roche, but was significant between VitaCyte and Serva (*p = 0.020)( Fig. 1B ). Digestion percentage showed significant difference between VitaCyte and Roche (*p = 0.026), as well as between VitaCyte and Serva (**p = 0.004) ( Fig. 1C ).

    Article Snippet: The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15).

    Techniques:

    Viability (A) and OCR (B) results of the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There is a significant difference in viability between VitaCyte and Roche (**p = 0.005), as well as between VitaCyte and Serva (*p = 0.020) ( Fig. 3A ). OCR data showed no significant difference between any two groups of enzymes ( Fig. 3B ).

    Journal: Islets

    Article Title: Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

    doi: 10.1080/19382014.2017.1417716

    Figure Lengend Snippet: Viability (A) and OCR (B) results of the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There is a significant difference in viability between VitaCyte and Roche (**p = 0.005), as well as between VitaCyte and Serva (*p = 0.020) ( Fig. 3A ). OCR data showed no significant difference between any two groups of enzymes ( Fig. 3B ).

    Article Snippet: The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15).

    Techniques:

    Purity post-purification (A) and packed cell volume post-purification (B) of the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There are no significant differences in both purity post-purification and packed cell volume post-purification between any two groups of enzymes.

    Journal: Islets

    Article Title: Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

    doi: 10.1080/19382014.2017.1417716

    Figure Lengend Snippet: Purity post-purification (A) and packed cell volume post-purification (B) of the three enzymes used. VitaCyte (n = 8), Roche (n = 48), and Serva (n = 15). There are no significant differences in both purity post-purification and packed cell volume post-purification between any two groups of enzymes.

    Article Snippet: The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15).

    Techniques: Purification

    Reversal rate of diabetes in NOD scid mice transplanted with islets isolated using three groups of enzymes. VitaCyte (n = 7), Roche (n = 34), and Serva (n = 13). No significant differences were found between VitaCyte enzyme group and any of the two other groups of enzymes. There was a significant difference between the Roche and Serva groups (**p = 0.008).

    Journal: Islets

    Article Title: Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

    doi: 10.1080/19382014.2017.1417716

    Figure Lengend Snippet: Reversal rate of diabetes in NOD scid mice transplanted with islets isolated using three groups of enzymes. VitaCyte (n = 7), Roche (n = 34), and Serva (n = 13). No significant differences were found between VitaCyte enzyme group and any of the two other groups of enzymes. There was a significant difference between the Roche and Serva groups (**p = 0.008).

    Article Snippet: The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15).

    Techniques: Mouse Assay, Isolation

    CD83’s TM domain is necessary and sufficient for CD4 T cell selection. (A) Experimental strategy to reconstitute expression of CD83 or chimeric variants thereof in TECs in vivo. WT or Cd83 −/− embryonic TECs, obtained by collagenase/dispase (C/D) digestion and CD45 depletion of E14–16 thymuses, were transduced with vectors encoding equimolar amounts of the construct of interest and EGFP via a T2A linker. RTOCs were prepared by 48-h cultivation on a nylon membrane and subsequently transplanted under the kidney capsule. Thymocyte subsets and TECs were analyzed 5–6 wk after transplantation. sinUTR, self-inactivating UTR; Ubi, ubiquitin; UTR, untranslated region. (B) Expression of the virally encoded GFP reporter in gated cTECs (CD45 − EpCAM + Ly51 + CD80 − ) and mTECs (CD45 − EpCAM + Ly51 − CD80 + ) from RTOCs prepared with a full-length CD83-encoding vector (#2 in F) on day 7 after transplantation. Gray histograms are noninfected control cTECs or mTECs. (C) GFP expression in TECs from RTOCs on days 5 and 35 after transplantation. Gray histograms are TECs from nontransduced RTOCs. (D) CD4 SP thymocyte frequencies in mixed RTOCs with titrated ratios of WT and Cd83 −/− TECs. Individual data points (one or two per time point) are indicated. (E) Thymocyte subsets in Cd83 −/− RTOCs or Cd83 −/− RTOCs from full-length CD83-transduced material. (F) Frequency of CD4 SP cells in RTOCs from Cd83 −/− TECs transduced with viral constructs (Constr.) #1–6 (schematically depicted below the respective data points). The mean transduction efficacy (percentage of GFP + cells among gated CD45 − EpCAM + TECs) ± SEM is shown. n ≥ 3. (G) TECs from RTOCs transduced with the indicated lentiviral constructs as indicated in F were stained for the EC domain of CD83. Unshaded histograms are gated on GFP + TECs (CD45 − EpCAM + cells). Gray histograms are gated on GFP − TECs. The MFI of gated GFP + TECs is indicated. Data in B, C, and E are representative of n ≥ 6 each in at least two independent experiments. In D and F, each data point corresponds to an individual RTOC. ***, P

    Journal: The Journal of Experimental Medicine

    Article Title: Thymic CD4 T cell selection requires attenuation of March8-mediated MHCII turnover in cortical epithelial cells through CD83

    doi: 10.1084/jem.20160316

    Figure Lengend Snippet: CD83’s TM domain is necessary and sufficient for CD4 T cell selection. (A) Experimental strategy to reconstitute expression of CD83 or chimeric variants thereof in TECs in vivo. WT or Cd83 −/− embryonic TECs, obtained by collagenase/dispase (C/D) digestion and CD45 depletion of E14–16 thymuses, were transduced with vectors encoding equimolar amounts of the construct of interest and EGFP via a T2A linker. RTOCs were prepared by 48-h cultivation on a nylon membrane and subsequently transplanted under the kidney capsule. Thymocyte subsets and TECs were analyzed 5–6 wk after transplantation. sinUTR, self-inactivating UTR; Ubi, ubiquitin; UTR, untranslated region. (B) Expression of the virally encoded GFP reporter in gated cTECs (CD45 − EpCAM + Ly51 + CD80 − ) and mTECs (CD45 − EpCAM + Ly51 − CD80 + ) from RTOCs prepared with a full-length CD83-encoding vector (#2 in F) on day 7 after transplantation. Gray histograms are noninfected control cTECs or mTECs. (C) GFP expression in TECs from RTOCs on days 5 and 35 after transplantation. Gray histograms are TECs from nontransduced RTOCs. (D) CD4 SP thymocyte frequencies in mixed RTOCs with titrated ratios of WT and Cd83 −/− TECs. Individual data points (one or two per time point) are indicated. (E) Thymocyte subsets in Cd83 −/− RTOCs or Cd83 −/− RTOCs from full-length CD83-transduced material. (F) Frequency of CD4 SP cells in RTOCs from Cd83 −/− TECs transduced with viral constructs (Constr.) #1–6 (schematically depicted below the respective data points). The mean transduction efficacy (percentage of GFP + cells among gated CD45 − EpCAM + TECs) ± SEM is shown. n ≥ 3. (G) TECs from RTOCs transduced with the indicated lentiviral constructs as indicated in F were stained for the EC domain of CD83. Unshaded histograms are gated on GFP + TECs (CD45 − EpCAM + cells). Gray histograms are gated on GFP − TECs. The MFI of gated GFP + TECs is indicated. Data in B, C, and E are representative of n ≥ 6 each in at least two independent experiments. In D and F, each data point corresponds to an individual RTOC. ***, P

    Article Snippet: RTOC Single cell suspensions of E14–E16 fetal thymic lobes were prepared by collagenase/dispase I (Roche) digestion.

    Techniques: Selection, Expressing, In Vivo, Transduction, Construct, Transplantation Assay, Plasmid Preparation, Staining

    Enriched pancreatic ducts and pancreatic duct epithelium in culture. A: Enriched pancreatic ducts after partial digestion with collagenase D and filtration through 85-μm nylon mesh. Phase contrast, original magnification, ×100. B: Primary culture of duct cells embedded in rat-tail collagen gel with a nylon mesh. Expanding duct epithelium can be seen in the lattice. Original magnification, ×100. C: Electron microscopy of the cultured pancreatic duct cells. Their epithelial nature was apparent by the prominent microvillus processes along portions of the plasma membrane ( thick white arrows ) and formed tight junctions ( solid black arrows ) between the cohered cells. No secretary granules are seen to suggest acinar cell derivation.

    Journal: The American Journal of Pathology

    Article Title: Liver Repopulation and Correction of Metabolic Liver Disease by Transplanted Adult Mouse Pancreatic Cells

    doi:

    Figure Lengend Snippet: Enriched pancreatic ducts and pancreatic duct epithelium in culture. A: Enriched pancreatic ducts after partial digestion with collagenase D and filtration through 85-μm nylon mesh. Phase contrast, original magnification, ×100. B: Primary culture of duct cells embedded in rat-tail collagen gel with a nylon mesh. Expanding duct epithelium can be seen in the lattice. Original magnification, ×100. C: Electron microscopy of the cultured pancreatic duct cells. Their epithelial nature was apparent by the prominent microvillus processes along portions of the plasma membrane ( thick white arrows ) and formed tight junctions ( solid black arrows ) between the cohered cells. No secretary granules are seen to suggest acinar cell derivation.

    Article Snippet: The harvested pancreas was immediately cut into small pieces and digested by collagenase D (Roche, Indianapolis, IN) (2.5 mg/ml, dissolved in Earle’s basic salt solution) for 25 minutes at 37°C with agitation by a magnetic stir bar.

    Techniques: Filtration, Electron Microscopy, Cell Culture