Collagenase type a

Manufactured by Merck Group

Sourced in United States

Collagenase type A is a laboratory reagent used for the enzymatic digestion of collagen. It is derived from Clostridium histolyticum and acts by cleaving the peptide bonds in collagen, facilitating the isolation and dissociation of cells from collagen-rich tissues.

Automatically generated - may contain errors

Lab products found in correlation

Hank s balanced salt solution, by Thermo Fisher Scientific (3 mentions) Ls006365, by Worthington (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Trypsin, by Merck Group (2 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Trypsin edta 0.05, by Thermo Fisher Scientific (1 mentions) Inhaled isoflurane anesthetic circuit 2, by Abbott (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Dnase, by Roche (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Penicillin streptomycin, by Lonza (1 mentions) L glutamine, by Lonza (1 mentions) 40 μm filter, by Corning (1 mentions) Hanks balanced salt solution (hbss), by Qiagen (1 mentions) Qiazol, by Qiagen (1 mentions) 48 well suspension culture plates, by Greiner (1 mentions) Falcon cell strainer, by BD (1 mentions) Falcon cell strainer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Collagenase A type I Collagenase A type I solution Collagenase crude type I A Type collagenase Collagenase A Type A Collagenase

17 protocols using collagenase type a

Quantifying Cell-Specific Drug Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

Apoe^−/−(Cdh5-CreER^T2;Apoe^−/−mT/mG mice treated
with Tamoxifen) mice were injected with 1 mg/kg 7C1 formulated with
Alexa647-tagged siLuciferase (VasoRx, Inc.). Two hours after intravenous
injection with 1 mg/kg 7C1-siLuciferase Alexa647, the aorta, heart, and lungs
were dissected, rinsed in cold PBS, and sliced into small pieces. The finely
minced tissue was transferred to a digestion mix consisting of Hank’s
balanced salt solution (Gibco) + 1 mg/ml collagenase type A (Sigma 10103578001)
+ 0.5 mg/ml elastase (Worthington LS006365) for 3 hr at 37°C and
pipetting every 30 min. The cell suspension was passed through a 40 μm
filter. DAPI (Sigma D9542) was used to detect dead cells. A FACS machine (BD
FACSAria) was used to analyze GFP⁺7C1-siLuciferase
Alexa647⁺ cells.

Chen P.Y., Qin L., Li G., Wang Z., Dahlman J.E., Malagon-Lopez J., Gujja S., Cilfone N.A., Kauffman K.J., Sun L., Sun H., Zhang X., Aryal B., Canfran-Duque A., Liu R., Kusters P., Sehgal A., Jiao Y., Anderson D.G., Gulcher J., Fernandez-Hernando C., Lutgens E., Schwartz M.A., Pober J.S., Chittenden T.W., Tellides G, & Simons M. (2019). Endothelial TGF-β signalling drives vascular inflammation and atherosclerosis. Nature metabolism, 1(9), 912-926.

+ Open protocol

+ Expand

Isolation and Culture of Cardiac Mesenchymal Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Green fluorescent protein (GFP) transgenic mice were deeply anesthetized by using an inhaled isoflurane anesthetic circuit 2% (Abbott, Abbott Park, IL, USA) for 10 minutes and euthanized by cervical dislocation. The hearts were then removed to isolate CMSCs. Cardiac fragments (approximately 1 mm) were dissected and incubated with 0.1% collagenase type A (Sigma-Aldrich, St. Louis, MO, USA) at 37°C for 30 minutes under constant stirring. After chemical and mechanical dissociation, fragments were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% Pen Strep (Gibco) as explants in sixwell plates and incubated at 37°C with 5% CO₂. Culture medium was changed every 3 days, and on day 8, upon reaching 90% confluence, the explants were removed from the wells and adherent cells were trypsinized (Trypsin-EDTA 0.05%; Gibco) and transferred to tissue culture flasks containing medium supplemented with 10% FBS and 1% Pen Strep. The CMSC culture was maintained in a humidified incubator at 37°C with 5% CO₂ for in vitro and in vivo studies.

Silva D.N., de Freitas Souza B.S., Azevedo C.M., Vasconcelos J.F., Carvalho R.H., Soares M.B, & dos Santos R.R. (2014). Intramyocardial transplantation of cardiac mesenchymal stem cells reduces myocarditis in a model of chronic Chagas disease cardiomyopathy. Stem Cell Research & Therapy, 5(4), 81.

+ Open protocol

+ Expand

Tumor Tissue Dissociation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Freshly obtained tumor material was dissociated or stored overnight in complete medium (RPMI-1640 with Hepes (Lonza, Verviers, Belgium), supplemented with 10% human serum (Sanquin, Amsterdam, The Netherlands), 1% L-glutamine (Lonza) and 1% penicillin/streptomycin (Lonza)) at 4°C prior to dissociation. The tumor tissue was weighed, cut in small fragments of approximately 1 mm³, placed in RPMI-1640 with Hepes supplemented with 1% penicillin/streptomycin, 0.1 mg/mL collagenase (Type A, Sigma-Aldrich, St. Louis, Missouri, USA) and 10 µg/mL DNAse (Roche), and mechanically dissociated using the gentleMACS Dissociator (program C and program h_tumor_01) (Miltenyi, Bergisch Gladbach, Germany). Subsequently, tumor pieces were incubated for 60 min at 37°C and 5% CO₂ under continuous rotation, after which cells were passed through a 70 µm strainer (Greiner Bio-One, Kremsmünster, Austria) and washed once with complete medium.

Klaver Y., Rijnders M., Oostvogels A., Wijers R., Smid M., Grünhagen D., Verhoef C., Sleijfer S., Lamers C, & Debets R. (2020). Differential quantities of immune checkpoint-expressing CD8 T cells in soft tissue sarcoma subtypes. Journal for Immunotherapy of Cancer, 8(2), e000271.

+ Open protocol

+ Expand

Kidney tubule isolation and RNA extraction

Check if the same lab product or an alternative is used in the 5 most similar protocols

Adult TuAB trunk kidneys (two per sample) were incubated at room temperature with 0.5mg/ml Collagenase Type A (Sigma, Saint Louis, MO) in HBSS (Walkersville, MD) for 30 minutes on a rocking stage at room temperature. The tissue was vigorously vortexed for 3 minutes to fragment the tissue. The tube was left to stand for 5 minutes to allow the tubules to separate from single cells by gravity. The supernatant was passed through a 40μm filter (Corning, Corning, NY), pelleted and lysed into Qiazol (Qiagen, Venlo, Netherlands) for RNA preparation. The tubular fraction was washed with HBSS and gravity collected again for 5 minutes, then lysed into Qiazol (Qiagen, Venlo, Netherlands) for RNA preparation. Enrichment for hematopoietic cell types in single cell fraction RNA and kidney tubular epithelial cells in tubule fraction RNA was confirmed by qRTPCR for gata1 (single cells) and cdh17 (tubule cells) (Supplemental figure 5).

Gallegos T.F., Kamei C.N., Rohly M, & Drummond I.A. (2019). Fibroblast growth factor signaling mediates progenitor cell aggregation and nephron regeneration in the adult zebrafish kidney. Developmental biology, 454(1), 44-51.

+ Open protocol

+ Expand

Intrahepatic Cholangiocyte Organoid Initiation

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Intrahepatic cholangiocyte organoids (ICO) (n = 5) were initiated from donor liver biopsies (0.5 cm³–1 cm³) following a previously published protocol [5 (link),22 (link)]. The use of liver biopsies for research purposes was approved by the medical ethics committee of the Erasmus University Medical Center (MEC-2014-060). Cholangiocarcinoma (CCA) tumor tissue biopsies (n = 3) were obtained through liver resections performed at the Erasmus MC (MEC-2013-143) and CCAO were initiated as described previously [9 (link)]. All patients gave written informed consent to use their tissue for research purposes. In short, the biopsies were digested in 2.5 mg/mL collagenase type A (Sigma-Aldrich, St. Louis, MO, US) for 20–120 min at 37 °C. Next, the suspension was strained (70 µm cell strainer), washed with cold ADV+ (Table S1), resuspended in basement membrane extract (BME, Cultrex, R&D systems, Minneapolis, MN, United States), and plated in 25 µL droplets in 48-well suspension culture plates (Greiner Bio One, Alphen aan den Rijn, The Netherlands). The BME was allowed to solidify for 45–60 min at 37 °C before 250 µL startup expansion medium (SEM, Table S2) was added. After 72 h, SEM was replaced with Expansion Medium (EM, Table S2) and refreshed every 3 to 4 days. Organoids were passaged by mechanical dissociation every 7 days in 1:3 to 1:6 split ratios.

van Tienderen G.S., Willemse J., van Loo B., van Hengel E.V., de Jonge J., van der Laan L.J., Leijten J, & Verstegen M.M. (2022). Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules. Cells, 11(22), 3657.

+ Open protocol

+ Expand

Quantifying Cell-Specific Drug Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Histological Evaluation of Murine Kidney

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The kidneys were fixed in 10% formalin overnight at 4°C, embedded in paraffin, and cut at 6 μM before being stained with H&E and periodic acid-Schiff reagent. We evaluated in a blinded fashion glomerular pathology (in 100 glomeruli/kidney) using a semi-quantitative scale as before (13 , 14 (link)).
Cells were extracted from murine spleens and lymph nodes by filtering the tissue through a 100-μm BD Biosciences Falcon cell strainer. The extracts were centrifuged at 1200 rpm for 5 min. ACK lysing buffer (Quality Biological) solution was added in the cell pellet to lyse the red cells. The treated cell pellet was subsequently washed once with DMEM cell culture medium and re-suspended in medium for further treatment or staining.
Murine kidney tissue was incubated with collagenase type A (10 μg/ml; Sigma-Aldrich) in cold Dulbecco’s PBS buffer with EDTA (2 μM) for 15 min at 37°C. The digested kidney tissue suspension was strained through a 100-μm BD Biosciences Falcon cell strainer (Fisher Scientific). The cells were centrifuged at 1200 rpm for 5 min and the cell pellet washed and re-suspended for treatment or staining.

Dai H., He F., Tsokos G.C, & Kyttaris V.C. (2017). IL-23 limits the production of IL-2 and promotes autoimmunity in lupus. Journal of immunology (Baltimore, Md. : 1950), 199(3), 903-910.

+ Open protocol

+ Expand

Isolation and Culture of Intrahepatic Cholangiocyte Organoids

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

As previously described,¹⁵ (link)^,¹⁶ (link) ICOs (n = 5) were initiated and cultured using an established protocol. In brief, donor liver tissue was minced, rinsed twice with Earle's balanced salt solution (EBSS, Thermo-Fisher, Waltham, USA), and digested in collagenase solution (2.5 mg/mL collagenase Type A, Sigma–Aldrich, St. Louis, USA) in Advanced DMEM/F-12 medium (Invitrogen, Carlsbad, USA) for 15 min at 37 °C. Cold Advanced DMEM/F-12 was added to stop the digestion. The single-cell suspension was filtered using a 70 μm Nylon cell strainer and centrifuged at 1500 rpm, 5 min, at 4 °C. After that, the cell pellets were collected at the bottom and subsequently mixed with BME (Basement Membrane Extract, Type 2, R&D Systems, Minneapolis, USA). Cell droplets were then seeded in multi-well plates. Initiating medium was added after the solidification of BME and the cells were cultured at 37 °C in a humidified atmosphere with 5% CO². After three days of culture in the initiating medium, the cells were refreshed with an expansion medium in which noggin, Wnt, Y27632, and hES cell cloning recovery supplement were deprived. Components supplemented in Advanced DMEM/F12 medium for initiating and expansion medium were listed in Table S2.

Shi S., Roest H.P., van den Bosch T.P., Bijvelds M.J., Boehnert M.U., de Jonge J., Dekker S.O., de Vries A.A., de Jonge H.R., Verstegen M.M, & van der Laan L.J. (2023). Modeling bile duct ischemia and reoxygenation injury in human cholangiocyte organoids for screening of novel cholangio-protective agents. eBioMedicine, 88, 104431.

+ Open protocol

+ Expand

Isolation of Aortic Smooth Muscle Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The ascending aorta of Apoe^−/− and TGFβR2^iSMC-Apoe mice were dissected from the mice and rinsed in cold PBS. The tissue was opened longitudinally and sliced into small fragments roughly 2 mm in length. The finely minced tissue was transferred to a digestion mix consisting of Hank’s balanced salt solution (Gibco) + 1 mg/ml collagenase type A (Sigma 10103578001) + 0.5 mg/ml elastase (Worthington LS006365) for 3 hrs at 37°C and pipetting every 30 min. DAPI (10 μg/ml; Sigma D9542) was used to detect dead cells. The cell suspension was passed through a 40 μm filter before sorting. A FACS machine (BD FACSAria) was used to sort GFP⁺RFP⁻ live cells. Single cells were sorted into 0.4% BSA/PBS.

Chen P.Y., Qin L., Li G., Malagon-Lopez J., Wang Z., Bergaya S., Gujja S., Caulk A.W., Murtada S.I., Zhang X., Zhuang Z.W., Rao D.A., Wang G., Tobiasova Z., Jiang B., Montgomery R.R., Sun L., Sun H., Fisher E.A., Gulcher J.R., Fernandez-Hernando C., Humphrey J.D., Tellides G., Chittenden T.W, & Simons M. (2020). Smooth muscle cell reprogramming in aortic aneurysms. Cell stem cell, 26(4), 542-557.e11.

+ Open protocol

+ Expand

Isolation and Culture of Murine and Human Tendon Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After euthanasia, tail tendons were dissected from mice, minced, and placed in alpha-modified Eagle’s medium (alpha-MEM; Gibco). Tissue was digested in 0.2% collagenase type A (Sigma-Aldrich) in phosphate-buffered saline for 3 hours. The digested tissue was passed through a 100-μm cell strainer, pelleted by centrifugation at 460g for 5 min; the supernatant was discarded; and the cells were resuspended and plated in supplemented culture medium [alpha-MEM with 10% fetal bovine serum (Sigma-Aldrich), 1% penicillin/streptomycin (Gibco), and 1% amphotericin B (Invitrogen)] at 37°C, 5% CO₂, and 95% humidity. The cells were used on passage 2. Human tendon-derived cells were extracted from biopsied hamstring tendon explants (age, 22 to 44 years). Cultures were maintained at 37°C in a humidified atmosphere of5% CO₂ for 28 days. Cells were subcultured and trypsinized at subconfluency and used at passage 3.

Abraham A.C., Shah S.A., Golman M., Song L., Li X., Kurtaliaj I., Akbar M., Millar N.L., Abu-Amer Y., Galatz L.M, & Thomopoulos S. (2019). Targeting the NF-κB signaling pathway in chronic tendon disease. Science translational medicine, 11(481), eaav4319.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!