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Collagenase type 2 solution

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Collagenase type II solution is a sterile-filtered enzyme preparation derived from Clostridium histolyticum. It is commonly used in cell isolation and tissue dissociation procedures.

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Lab products found in correlation

Advanced dmem f12, by Thermo Fisher Scientific (2 mentions) Y 27632, by Merck Group (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) N acetyl l cysteine, by Merck Group (2 mentions) A83 01, by Bio-Techne (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Y 27632 dihydrochloride, by AbMole (1 mentions)

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Collagenase type II (621 citations) Collagenase (527 citations) Collagenase II (403 citations) Collagenase solution (11 citations) Collagenase II solution (9 citations) Type II (4 citations)

6 protocols using collagenase type 2 solution

1

Prostate Organoid Culture Protocol

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Prostate organoid culture was performed as described previously.[65] Briefly, minced prostate tissue from 8 to 12 weeks old male mice was digested in 5 mg mL−1 collagenase type II solution (Life Technologies, cat. no. 17101‐015) with 10 × 10−6m Y‐27632 dihydrochloride (Abmole Bioscience, cat. no. M1817) on a shaking platform at 37 °C for 1–1.5 h. After aspirating, the collagenase type II solution, a single prostate epithelial cell suspension was generated by incubating samples in TrypLE with 10 × 10−6m Y‐27632 dihydrochloride at 37 °C for 15 min. 20 000 prostate epithelial cells were embedded in a 40 µL Matrigel or PIC hydrogel drop and seeded in the middle of one well of a 24‐well tissue culture plate. After gelation of the Matrigel or PIC hydrogel, 500 µL prewarmed 3D prostate organoid culture medium (Table S1, Supporting Information) was added and organoids were cultured in an incubator (5% CO2, 37 °C). Culture medium was refreshed every 2–3 days.
Zhang Y., Tang C., Span P.N., Rowan A.E., Aalders T.W., Schalken J.A., Adema G.J., Kouwer P.H., Zegers M.M, & Ansems M. (2020). Polyisocyanide Hydrogels as a Tunable Platform for Mammary Gland Organoid Formation. Advanced Science, 7(18), 2001797.
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2

Bladder Organoid Culture Protocol

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Organoid lines were generated either from normal murine bladder or bladder tumors. Minced tissues were treated with 5 mg/mL collagenase type II solution (Life Technologies, 17101015) diluted in Dulbecco’s Modified Eagle’s Medium for 1 hour at 37°C, followed by a 5 minute TrypLE (Gibco, 12604–039) treatment at 37°C. Digested tissues further dissociated using a syringe with 18G needle, and single cells were obtained after passing through a cell strainer. Cells were plated using EHS (NIH and Corning) and cultured in organoid media for 5–7 days at 37°C. Organoid base media was prepared using Advanced DMEM/F12 (Gibco 12634010) supplemented with 20% B27 (Gibco, 17504–044), 10 mM HEPES, Glutamax (Fisher, 35050061), and 1.25 mM N-Acetyl-L-cysteine (Sigma, A9165). Base media was also supplemented with 50 ng/mL EGF (Peprotech, 315–09), 100 ng/mL Noggin (conditioned media), 500 ng/mL R-spondin (conditioned media), 200 nM A83–01 (Tocris, 2939), and 10 μM Y-27632 (Sigma, Y0503). Organoids were passaged once every week.
Jana S., Brahma S., Arora S., Wladyka C.L., Hoang P., Blinka S., Hough R., Horn J.L., Liu Y., Wang L.J., Depeille P., Smith E., Montgomery R.B., Lee J.K., Haffner M.C., Vakar-Lopez F., Grivas P., Wright J.L., Lam H.M., Black P.C., Roose J.P., Ryazanov A.G., Subramaniam A.R., Henikoff S, & Hsieh A.C. (2023). Transcriptional-translational conflict is a barrier to cellular transformation and cancer progression. Cancer cell, 41(5), 853-870.e13.
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3

Bladder Organoid Culture Protocol

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Organoid lines were generated either from normal murine bladder or bladder tumors. Minced tissues were treated with 5 mg/mL collagenase type II solution (Life Technologies, 17101015) diluted in Dulbecco’s Modified Eagle’s Medium for 1 hour at 37°C, followed by a 5 minute TrypLE (Gibco, 12604–039) treatment at 37°C. Digested tissues further dissociated using a syringe with 18G needle, and single cells were obtained after passing through a cell strainer. Cells were plated using EHS (NIH and Corning) and cultured in organoid media for 5–7 days at 37°C. Organoid base media was prepared using Advanced DMEM/F12 (Gibco 12634010) supplemented with 20% B27 (Gibco, 17504–044), 10 mM HEPES, Glutamax (Fisher, 35050061), and 1.25 mM N-Acetyl-L-cysteine (Sigma, A9165). Base media was also supplemented with 50 ng/mL EGF (Peprotech, 315–09), 100 ng/mL Noggin (conditioned media), 500 ng/mL R-spondin (conditioned media), 200 nM A83–01 (Tocris, 2939), and 10 μM Y-27632 (Sigma, Y0503). Organoids were passaged once every week.
Jana S., Brahma S., Arora S., Wladyka C.L., Hoang P., Blinka S., Hough R., Horn J.L., Liu Y., Wang L.J., Depeille P., Smith E., Montgomery R.B., Lee J.K., Haffner M.C., Vakar-Lopez F., Grivas P., Wright J.L., Lam H.M., Black P.C., Roose J.P., Ryazanov A.G., Subramaniam A.R., Henikoff S, & Hsieh A.C. (2023). Transcriptional-translational conflict is a barrier to cellular transformation and cancer progression. Cancer cell, 41(5), 853-870.e13.
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4

Isolation and Expansion of Human Adipose-Derived Stem Cells

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Human adipose tissue was obtained from the abdominal fat of a female subjected to cosmetic liposuction who previously signed an informed consent regarding donation of her fat for research. The protocol was approved by the Research and Ethical Committee of CUCS, University of Guadalajara (approval number C.I. 005–2017) which approved the fat obtainment procedure. Tissue was digested by a 0.075% collagenase type II solution (Invitrogen, Grand Island, NY) gently shaken for 1 hour at 37°C. Digestion product was filtered using a 100 um nylon mesh and centrifuged at 1200 g for 8 min. Pellet was washed with PBS and erythrocytes were lysed. Cells were collected and plated on plastic dishes in DMEM (Invitrogen, Grand Island, NY) supplemented with 10% fetal bovine serum (Invitrogen, Grand Island, NY) and 1% antibiotic (Invitrogen, Grand Island, NY). The medium was changed after 48 h. Cells were harvested and seeded until passage 3 to achieve greater expansion. Cell characterization and transplantation were made in this unique batch of cells obtained from one single fat donor in a sole isolation.
Rivera-Valdés J.J., García-Bañuelos J., Salazar-Montes A., García-Benavides L., Rosales-Dominguez A., Armendáriz-Borunda J, & Sandoval-Rodríguez A. (2017). Human adipose derived stem cells regress fibrosis in a chronic renal fibrotic model induced by adenine. PLoS ONE, 12(12), e0187907.
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5

Isolation of Neonatal Mouse Cardiomyocytes

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Cardiomyocytes were isolated from ventricles of C57BL/6J and C57BL/6N as described previously [13 (link)]. Briefly, cardiac myocytes were harvested from 1-day-old neonatal mice with the digestion of collagenase type II solution (1mg/ml, Invitrogen).The cells were cultured for 48 hours and collected for experiments.
Taniguchi M., Okamoto R., Ito M., Goto I., Fujita S., Konishi K., Mizutani H., Dohi K., Hartshorne D.J, & Itoh T. (2015). New Isoform of Cardiac Myosin Light Chain Kinase and the Role of Cardiac Myosin Phosphorylation in α1-Adrenoceptor Mediated Inotropic Response. PLoS ONE, 10(10), e0141130.
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6

Isolation of Human Adipose-Derived Cells

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Human adipose tissue was obtained from abdominal fat of females subjected to cosmetic liposuction who signed a written consent to donate the discarded fat tissue. The protocol was approved by the Research and Ethical Committee of the CUCS, Universidad de Guadalajara (approval number C.I. 067–2012) which review the fat obtainment procedure. Tissue was digested by a 0.075% collagenase type II solution (Invitrogen, Grand Island, NY) for 1 hour at 37°C with gentle shaking. Digestion product was filtered using a 100 um nylon mesh and centrifuged at 1200 g for 8 min. Pellet was washed with PBS once. Cells were collected and plated on plastic dishes in DMEM (Invitrogen, Grand Island, NY) supplemented with 10% fetal bovine serum (Invitrogen, Grand Island, NY) and 1% antibiotic (Invitrogen, Grand Island, NY). The medium was changed after 48 h. Cells were harvested and seeded until passage three for achieving greater expansion. Cell characterization and transplantation was made using a unique batch of cells from a sole isolation.
Meza-Ríos A., García-Benavides L., García-Bañuelos J., Salazar-Montes A., Armendáriz-Borunda J, & Sandoval-Rodríguez A. (2016). Simultaneous Administration of ADSCs-Based Therapy and Gene Therapy Using Ad-huPA Reduces Experimental Liver Fibrosis. PLoS ONE, 11(12), e0166849.
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