Collagenase Perfusion
Collagenase Perfusion - 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.
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.
06:59
Mouse Islet of Langerhans Isolation using a Combination of Purified Collagenase and Neutral Protease
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Cited by 103 •
2012
A detailed description of mouse islet isolation is described using the technique of in situ pancreatic ductal cannulation and perfusion of a combination of purified collagenase and neutral protease.
FAQs
Collagenase cleaves collagen in the extracellular matrix, weakening the structural framework that helps retain cells within tissue. Vascular delivery distributes the buffered enzyme solution through the organ, promoting exposure beyond the tissue surface. This combination loosens cellular connections while limiting the need for forceful mechanical disruption, which can help preserve cells for subsequent analysis.
Perfusion uses the organ’s vascular pathways to circulate collagenase through intact tissue, rather than relying primarily on contact at an external surface. More even enzyme exposure can improve the release of cells from complex internal regions. The resulting dissociation is particularly useful when investigators need to examine immune or tissue-resident populations that might be unevenly recovered by surface treatment.
The buffered collagenase solution provides the enzyme in a circulating medium suitable for delivery through the tissue. Collagenase supplies the matrix-cleaving activity, while circulation supports distribution throughout the organ. Together, these features promote loosening of cellular connections and help produce a cell suspension appropriate for downstream flow cytometry, culture, microscopy, or molecular assays.
By reducing dependence on vigorous mechanical disruption, collagenase perfusion can support recovery of viable immune and tissue-resident cells from complex organs. Preserving these cells matters because downstream measurements may depend on their intact characteristics and interactions. In immunology, improved recovery can strengthen analyses of inflammatory responses and cellular composition within tissue rather than only in readily accessible samples.
A basic workflow includes preparing a buffered collagenase solution, delivering it through the organ’s vasculature, allowing circulation to expose internal tissue regions, and collecting the released cells for analysis. The recovered material can then be directed toward flow cytometry, culture, microscopy, or molecular assays. The central procedural goal is thorough exposure with minimal mechanical damage.
Recovered cells can support several complementary readouts. Flow cytometry can characterize cellular populations, culture can examine cell behavior, microscopy can assess cells or tissue-associated features, and molecular assays can investigate biological changes. Using the same dissociation approach across these applications can help connect cellular composition with functional, visual, or molecular observations.
The method helps investigators recover immune and tissue-resident cells from intact organs, where inflammatory or pathogen-associated changes may be distributed across complex tissue environments. Those cells can be examined using cellular, microscopic, culture-based, or molecular approaches. This makes the technique useful for studying inflammatory responses, pathogen-associated alterations, and interactions between immune cells and tissue compartments.