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Collagenase elastase

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Collagenase/Elastase is a lab equipment product that functions to break down collagen and elastin, which are structural components of connective tissue. It is a mixture of enzymes used for tissue dissociation and cell isolation applications.

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Lab products found in correlation

Liver perfusion media, by Thermo Fisher Scientific (2 mentions) Dnase 1, by Worthington (2 mentions) Deoxyribonuclease, by Worthington (1 mentions) Amino acids, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions) Hank s buffered salt solution, by Merck Group (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)

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Collagenase (413 citations) Elastase (132 citations)

3 protocols using collagenase elastase

1

Isolation of Primary Hepatocytes

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Primary hepatocytes were isolated as previously described (41 (link)) using a modified 2-step perfusion method. The portal vein was cannulated, and the liver was perfused with liver perfusion media (Invitrogen, 17701-038) followed by liver digestion media, containing Krebs-Ringer Bicarbonate (KRB, MilliporeSigma, K-4002) supplemented with 20 mM HEPES, 500 μM CaCl2, Collagenase/Elastase (Worthington, LK002066) and DNase I (Worthington, LK003170). After perfusion, the liver was removed and disrupted to release cells using cell scrapers. The cell suspension was then filtered through a 70 μm filter, centrifuged at 50g for 5 minutes at 4°C, washed once in Krebs-Ringer buffer, and precipitated in a 25% Percoll gradient at 120g for 5 minutes at 4°C. NR treatment was 500 μM, which we have found to be optimal for increasing NAD content.
Mukherjee S., Mo J., Paolella L.M., Perry C.E., Toth J., Hugo M.M., Chu Q., Tong Q., Chellappa K, & Baur J.A. (2021). SIRT3 is required for liver regeneration but not for the beneficial effect of nicotinamide riboside. JCI Insight, 6(7), e147193.
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2

Cell Culture Media Composition

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Collagenase/elastase and deoxyribonuclease were purchased from Worthington Biochemical Corporation (Lakewood NJ). Cell culture reagents, including Minimal Essential Media alpha, fetal bovine serum, antibiotic-antimycotic solution and amino acids, were purchased from Life Technologies (Carlsbad CA). Hank’s buffered salt solution and other chemicals were acquired from Sigma (St Louis MO). Epidermal growth factor (EGF) and porcine insulin were purchased from PeproTech (Rocky Hill NJ).
Gruppuso P.A., Boylan J.M., Zabala V., Neretti N., Abshiru N.A., Sikora J.W., Doud E.H., Camarillo J.M., Thomas P.M., Kelleher N.L, & Sanders J.A. (2018). Stability of histone post-translational modifications in samples derived from liver tissue and primary hepatic cells. PLoS ONE, 13(9), e0203351.
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3

Isolation of Primary Murine Hepatocytes

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Primary hepatocytes (male) were isolated from 8-weeks-old male C57BL/6J mice, ordered directly from Jackson Labs. Animals were housed in groups of five mice/cage in a pathogen-free barrier facility in a 12-hour light-dark cycle with free access to food and water for 2weeks.
Hepatocytes were isolated using modified two-step perfusion method. The portal vein was cannulated and the liver was perfused with Liver Perfusion Media (Invitrogen) followed by Liver Digestion media, containing Kreb-Ringer Bicarbonate (Sigma) supplemented with 20mM HEPES, 500uM CaCl2, Collagenase/Elastase (Worthington) and DNase I (Worthington) were used for isolation. After the perfusion, liver was removed, disrupted to release cells using cell scrapers. The cell suspension was then filtered through 70μm filter and centrifuged at 50g for 5 minutes at 4°C, washed once in KRB buffer and precipitated in 25% Percoll gradient at 120g for 5 minutes at 4°C. Healthy hepatocytes were plated at a concentration of 3X106 and 1X106 on 10 cm and 100 mm collagen coated dishes respectively, in 1XDMEM supplemented with 10% FBS and1% Penicillin Streptomycin (Miller, et al, JCI, 2011 ).
Krishnaiah S.Y., Wu G., Altman B.J., Growe J., Rhoades S.D., Coldren F., Venkataraman A., Olarerin-George A.O., Francey L.J., Mukherjee S., Girish S., Selby C.P., Cal S., Ubeydullah E., Sianati B., Sengupta A., Anafi R.C., Kavakli I.H., Sancar A., Baur J.A., Dang C.V., Hogenesch J.B, & Weljie A.M. (2017). Clock regulation of metabolites reveals coupling between transcription and metabolism. Cell metabolism, 25(4), 961-974.e4.
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