Collagenase Digestion
Collagenase Digestion - Related Videos
07:09
Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure
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Cited by 93 •
2013
A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.
07:21
Pancreatic Islets Isolation through Collagenase Perfusion: An Enzymatic Method to Isolate Pancreatic Islet Cells
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2023
Pancreatic islets, also called the Islets of Langerhans, are a cluster of endocrine cells which produces hormones for glucose regulation and other important biological functions. This protocol explains a method for isolating pancreatic islets by first perfusing collagenase via Ampulla of Vater to digest the exocrine pancreatic tissue and a simplified density gradient centrifugation to purify the islets from mice.
09:41
Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage
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Cited by 35 •
2014
Preclinical models of intracerebral hemorrhage are utilized to mimic certain aspects of clinical disease. Thus, mechanisms of injury and potential therapeutic strategies may be explored. In this protocol, two models of intracerebral hemorrhage are described, intrastriatal (basal ganglia) injections of autologous blood or collagenase.
09:45
Restriction Enzyme Digests
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2023
Restriction enzymes or endonucleases recognize and cut DNA at a specific sequence. These enzymes occur naturally in bacteria as a defense against bacteriophages - viruses that infect bacteria. Bacterial restriction enzymes cut the invading bacteriophage DNA while leaving the bacterial genomic DNA unharmed due to addition of methyl groups. This video explains the basic principles of restriction enzymes including: how restriction enzymes are named and the types of recognition sites and...
10:44
Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase
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Cited by 60 •
2012
Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.
FAQs
The enzyme cleaves collagen fibers within connective tissue, weakening the matrix that holds cells and other components in place. This loss of structural support allows embedded material to separate from the tissue through enzymatic treatment rather than mechanical disruption alone. The extent of cleavage therefore affects how efficiently cells can be released for subsequent culture or analysis.
Digestion conditions determine whether the matrix is sufficiently loosened while the recovered cells remain suitable for study. Buffer composition, temperature, and exposure time are specifically important because the process is performed in buffered solutions under controlled temperature and timing. Excessive or insufficient treatment can alter the balance between tissue dissociation and preservation of the cellular material needed for downstream work.
Collagenase targets the collagen-rich structural framework, whereas supporting enzymes can assist with additional extracellular matrix proteins. Their complementary action may improve access to cells embedded in a complex tissue rather than relying on collagen cleavage alone. The choice to include supporting activity depends on the matrix components that need to be dissociated and on whether released cells or other matrix material is the desired outcome.
Preserving cell viability and surface markers is a central constraint, not merely a technical preference. Digestion conditions must be controlled so that cells are released without compromising their ability to remain viable or retain identifying features on their surfaces. This matters when isolated primary cells are subsequently cultured or analyzed, because preparation-related changes could affect interpretation of their biological properties.
A basic workflow places tissue in a buffered collagenase solution, maintains controlled temperature and exposure time, and then proceeds with the released cells or matrix components for culture or analysis. The treatment must be carefully controlled for the tissue being processed because skin, adipose tissue, pancreas, and tumors differ in their connective-tissue context and intended experimental outcome.
Collagenase digestion supports isolation of primary cells from skin, adipose tissue, pancreas, and tumors. Once released, these cells can support culture and analysis, while accompanying matrix components may also be examined. The method therefore connects tissue processing with studies of tissue engineering, disease, organoids, and regenerative medicine, where access to cells from their original tissue environment is important.