Collagenase 1

Manufactured by Fujifilm

Sourced in Japan

Collagenase I is an enzyme used in cell culture and tissue engineering applications. It is primarily responsible for the digestion of collagen, a structural protein found in the extracellular matrix of various tissues. Collagenase I is commonly employed in the isolation and dissociation of cells from tissues, enabling researchers to obtain individual cells for further experimentation or cultivation.

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Lab products found in correlation

Facs calibur hg flow cytometer, by BD (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Cellquest software, by BD (2 mentions) Dnase 1, by Fujifilm (2 mentions) Clone fjk 16s, by Abcam (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Dispase 2, by Fujifilm (2 mentions) Facsverse, by BD (1 mentions) Trastuzumab, by Roche (1 mentions) Herceptin, by Roche (1 mentions) Anti cd16 32 2.4g2, by BD (1 mentions) I a 1 e, by BD (1 mentions) Cd11b, by BioLegend (1 mentions) F4 80, by BioLegend (1 mentions) Dulbecco modified eagle medium dmem, by Merck Group (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Fungizone, by Thermo Fisher Scientific (1 mentions) L ascorbic acid 2 phosphate, by Fujifilm (1 mentions)

Spelling variants of this product

Collagenase (247 citations) Collagenase type I (58 citations)

9 protocols using collagenase 1

Tumor and Lymph Node Cell Isolation

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Fresh tumor and metastatic lymph node tissues were minced with scalpels and digested in 1 mg/ml collagenase I (Wako, Osaka, Japan) with 0.5% bovine serum albumin at 37°C
for 2 hr. Undigested tissue was removed by filtration using a 100 µm mesh. Cells were incubated with 200 µl of 10 µg/mltrastuzumab (Herceptin; Roche, Hertfordshire, U.K.) or without trastuzumab as negative control for 30 min at 4°C, followed by three washes in fluorescence-activated cell sorting (FACS)
buffer (phosphate-buffered saline containing 5% fetal bovine serum). Then, cells were incubated with anti-human IgG APC antibodies (BioLegend) diluted 1:25 for 30 min at 4°C, and washed with
FACS buffer. Negative controls were also subjected to the same procedures. After live cells were gated by forward scatter/side scatter (FSC/SSC), fluorescence was measured and analyzed using
a flow cytometer and its related software (BD FACSVerse™, Becton-Dickinson, Franklin Lakes, NJ, U.S.A.). Staining positivity was confirmed by negative control.

YOSHIMOTO S., KATO D., KAMOTO S., YAMAMOTO K., TSUBOI M., SHINADA M., IKEDA N., TANAKA Y., YOSHITAKE R., ETO S., SAEKI K., CHAMBERS J.K., KINOSHITA R., UCHIDA K., NISHIMURA R, & NAKAGAWA T. (2019). Detection of human epidermal growth factor receptor 2 overexpression in canine anal sac gland carcinoma. The Journal of Veterinary Medical Science, 81(7), 1034-1039.

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Phenotypic Analysis of Tumor Immune Cells

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Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). The resulting cell suspensions were clarified using 40-μm filters to prepare single cell suspensions, and single cells were suspended in PBS supplemented with 2% FBS. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 (BD Biosciences, San Jose, CA, USA) to reduce FcγR binding. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, USA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), CD45 (BioLegend, clone 30-F11), IA/IE (BD Bioscience, clone M5/114), CD11c (BD Bioscience, clone HL3), CD11b (BioLegend, clone M1/70), and F4/80 (BioLegend, clone BM8).
For intracellular staining, cells were fixed and permeabilized using Foxp3/Transcription Factor Staining Buffer Set (eBioscience, San Diego, CA, USA) after cell surface staining and were stained with labeled antibodies against the intracellular molecules foxp3 (eBioscience, clone FJK-16s), glucose transporter 1 (GLUT1, Abcam, Cambridge, UK, clone EPR3915), and hexokinase II (HX2, Abcam, clone EPR20839).
Samples were analyzed on a FACS Calibur HG flow cytometer (BD Biosciences). Data analysis was performed with CellQuest™ software (Becton Dickinson, Lincoln Park, NJ, USA).

Tomita M., Yasui H., Higashikawa K., Nakajima K., Takakura H., Shiga T., Kuge Y, & Ogawa M. (2018). Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model. EJNMMI Research, 8, 82.

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Isolation and Culture of Mature Adipocytes

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sWAT was excised from euthanized adult male mice, minced with scissors, and incubated in collagenase buffer (20 mM HEPES (pH 7.4), 120 mM NaCl, 5 mM KCl, 4 mM NaHCO₃, 1 mM CaCl₂, 0.7 mM MgSO₄, 0.4 mM KH₂PO₄, 0.3 mM Na₂HPO₄, and 4 mg/mL collagenase I (031-17601, Fujifilm Wako, Osaka, Japan)) at 37 °C for 1 h. Subsequently, the floating layer containing mature adipocytes was collected. Isolated mature adipocytes were then resuspended and cultivated in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% (v/v) fetal bovine serum, 1 mM sodium pyruvate, 500 IU/mL penicillin, and 500 μg/mL streptomycin.

Hasegawa K., Takenaka N., Tanida K., Chan M.P., Sakata M., Aiba A, & Satoh T. (2021). Atrophy of White Adipose Tissue Accompanied with Decreased Insulin-Stimulated Glucose Uptake in Mice Lacking the Small GTPase Rac1 Specifically in Adipocytes. International Journal of Molecular Sciences, 22(19), 10753.

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Isolation and Expansion of Rat Adipose-Derived Stem Cells

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Under general anesthesia with 3 mixed anesthetics (midazolam 5 mg/kg, medetomidine 0.375 mg/kg, butorphanol 2.5 mg/kg), ASCs were isolated from the inguinal fat pads of Sprague–Dawley rat (8 weeks old; Japan SLC Inc.), according to the method previously reported [29 ]. In brief, subcutaneous adipose tissue was digested with 0.25 w/v% collagenase I (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) under gentle agitation for 45 min at 37 °C and added Dulbecco modified Eagle medium (DMEM, Sigma Aldrich, Missouri, United States) containing 10 v/v% fetal bovine serum (FBS, Sigma Aldrich) and centrifuged at 200 g for 5 min. After aspirating to remove cellular debris and supernatant, the pellet was suspended in the control medium (DMEM supplemented with 10 v/v% FBS and 1 v/v% antibiotic/antimycotic). Next, the cell suspension was filtered through a 40 μm nylon sieve to remove cellular debris and centrifuged at 200 g for 5 min. Then, the pellet was resuspended in a control medium and cultured in 100 mm tissue culture dishes at a density of 1 × 10⁶ cells per plate. These cells were cultured in the control medium at 37 °C and in 5% CO₂. ASCs at passage 3 isolated from rats were used for this study.

Mitsui R., Matsukawa M., Nakagawa K., Isomura E., Kuwahara T., Nii T., Tanaka S, & Tabata Y. (2021). Efficient cell transplantation combining injectable hydrogels with control release of growth factors. Regenerative Therapy, 18, 372-383.

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Isolation of Deciduous Tooth Stem Cells

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Human exfoliated deciduous teeth were provided by Pediatric Dentistry and Special Need Dentistry at Kyushu University Hospital in Japan. After informed parental consent was obtained, deciduous teeth were collected from a normal participant and a patient with DS at 6 and 14 years of age, respectively. The isolation procedure was completed as previously described [15 (link)]. Briefly, the pulp tissue was subjected to an enzymatic dissociation in 3 mg/mL collagenase I (Washington, NJ, USA) and 4 mg/mL dispase II (Wako, Osaka, Japan) for 1 h, and then maintained at 37 °C in a humidified 5% CO₂ incubator in the Alpha modification of Eagle’s Minimal Essential Medium (α-MEM; Sigma-Aldrich, MO, USA) containing 15% fetal bovine serum (Sigma-Aldrich), 100 μM L-ascorbic acid 2-phosphate (Wako), 2 mM L-glutamine (Life Technologies, NY, USA), 250 μg/mL Fungizone (Life Technologies), 100 U/mL penicillin (Life Technologies), and 100 μg/mL streptomycin (Life Technologies). Cells of not more than 10 passages were used, but Ctrl-SHED and DS-SHED were not always of the same passage.

Pham T.T., Kato H., Yamaza H., Masuda K., Hirofuji Y., Sato H., Nguyen H.T., Han X., Zhang Y., Taguchi T, & Nonaka K. (2018). Altered development of dopaminergic neurons differentiated from stem cells from human exfoliated deciduous teeth of a patient with Down syndrome. BMC Neurology, 18, 132.

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Immune Profiling of Tumor Microenvironment

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Thirteen mice were used for flow-cytometry (cGAMP alone group n = 6, cGAMP/anti-PD-1 combination group n = 7). Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). The resulting cell suspensions were clarified using 40-μm filters to prepare single cell suspensions, and single cells were suspended in PBS supplemented with 2% FBS. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 antibody (BD Biosciences, San Jose, CA) to reduce FcγR binding. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), and CD45 (BioLegend, clone 30-F11).
For intracellular staining, cells were fixed and permeabilized using a Foxp3/Transcription Factor Staining Buffer Set (eBioscience, San Diego, CA) after cell surface staining and then stained with labeled antibodies against the intracellular molecules Foxp3 (eBioscience, clone FJK-16s), glucose transporter 1 (GLUT1, Abcam, Cambridge, UK, clone EPR3915), and hexokinase II (HX2, Abcam, clone EPR20839).
Samples were analyzed on a FACS Calibur HG flow cytometer (BD Biosciences). Data analysis was performed with CellQuest™ software (Becton Dickinson, Lincoln Park, NJ).

Tomita M., Suzuki M., Kono Y., Nakajima K., Matsuda T., Kuge Y, & Ogawa M. (2020). Influence on [18F]FDG uptake by cancer cells after anti-PD-1 therapy in an enforced-immune activated mouse tumor. EJNMMI Research, 10, 24.

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Isolation of CD31-positive Endothelial Cells

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CD31-positive ECs were isolated from the WAT (epididymal fat pad) by the use of a Magnetic Cell Sorting System (MACS, Miltenyi Biotec, Auburn, CA). Tissues of WT or Vash1^−/− mice were minced and digested with collagenase I and Dispase II (Wako). The cell suspensions were then filtered through a 70 μm cell strainer (BD) and treated with ACK Lysing Buffer (GIBCO) to remove erythrocytes. CD31-positive ECs were isolated by MACS with CD31 MicroBeads (Miltenyi Biotec) according to the manufacturer's instructions.

Takeda E., Suzuki Y., Yamada T., Katagiri H, & Sato Y. (2017). Knockout of Vasohibin-1 Gene in Mice Results in Healthy Longevity with Reduced Expression of Insulin Receptor, Insulin Receptor Substrate 1, and Insulin Receptor Substrate 2 in Their White Adipose Tissue. Journal of Aging Research, 2017, 9851380.

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Isolation and Culture of Murine Adipose Stromal Cells

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Subcutaneous WAT was excised from euthanized 22-week-old male mice. Minced subcutaneous WAT was digested in collagenase buffer (20 mM HEPES (pH 7.4), 120 mM NaCl, 5 mM KCl, 4 mM NaHCO₃, 1 mM CaCl₂, 0.7 mM MgSO₄, 0.4 mM KH₂PO₄, and 0.3 mM Na₂HPO₄) supplemented with 3 mg/mL collagenase I (031-17601, Fujifilm Wako (Osaka, Japan)) at 37 °C for 1 h. Digested subcutaneous WAT was filtered through 100 μm nylon mesh to get a single-cell suspension, which was then centrifugated at 760× g for 10 min at room temperature. The precipitated SVF was suspended in KBM ADSC-1 (Kohjin Bio, Saitama, Japan) supplemented with 2500 IU/mL penicillin and 2500 μg/mL streptomycin and cultured at 37 °C with 5% CO₂ to confluence.

Hasegawa K., Takenaka N., Yamamoto M., Sakoda Y., Aiba A, & Satoh T. (2023). Regulation of De Novo Lipid Synthesis by the Small GTPase Rac1 in the Adipogenic Differentiation of Progenitor Cells from Mouse White Adipose Tissue. International Journal of Molecular Sciences, 24(5), 4608.

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Osteoblast Culture from Mouse Calvaria

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Primary osteoblasts were prepared from calvariae of 3-5-dayold C57BL/6 mice by five sequential digestions using 0.1% collagenase I (Wako Pure Chemicals). The cells were grown for 21 days in α-MEM containing 10% FBS in the presence of β-glycerophosphate and ascorbic acid (Wako Pure Chemicals). Alkaline phosphatase (ALP) activity was examined using TRAP/ALP Staining kit (Wako Pure Chemicals) according to the protocol provided in the kit. For 1,25(OH) 2 D 3 or PTH stimulation, primary osteoblasts were treated with 10 nM of 1,25(OH) 2 D 3 (Cayman, Ann Arbor, MI, USA) or 100 ng/ml of human PTH (1-31) (Bachem Bioscience, King of Prussia, PA, USA) as indicated in each experiment.

Uehara N., Kukita A., Kyumoto-Nakamura Y., Yamaza T., Yasuda H, & Kukita T. (2017). Osteoblast-derived Laminin-332 is a novel negative regulator of osteoclastogenesis in bone microenvironments. Laboratory investigation; a journal of technical methods and pathology, 97(10).

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