Lamina Propria Digestion
Lamina Propria Digestion - Related Videos
04:52
Isolating Enteric Glial Cells From the Submucosa and Lamina Propria of a Mouse
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2025
This video demonstrates a method for isolating enteric glial cells from the submucosa and lamina propria of the mouse intestine. It outlines the steps involved in extracting submucosa and lamina propria tissue, isolating the enteric glial cells from them, and culturing the cells on a coated well-plate to establish an enteric glial cell culture.
05:01
Lamina Propria Mononuclear Cell Isolation: A Silica-based Density Gradient Centrifugation Technique to Obtain Mononuclear Cells from Murine Colon
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2023
In this video, we demonstrate the isolation of lamina propria mononuclear cells from murine colonic tissue by enzymatic digestion using collagenase E and silica-based density separation media gradient separation. The isolated mononuclear cells can be used for subsequent downstream analysis.
09:48
Isolation of Lamina Propria Mononuclear Cells from Murine Colon Using Collagenase E
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Cited by 9 •
2019
The goal of this protocol is to isolate mononuclear cells that reside in the lamina propria of the colon by enzymatic digestion of the tissue using collagenase. This protocol allows for the efficient isolation of mononuclear cells resulting in a single cell suspension which in turn can be used for robust immunophenotyping.
07:53
Isolation of Enteric Glial Cells from the Submucosa and Lamina Propria of the Adult Mouse
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Cited by 21 •
2018
Here, we describe the isolation of enteric-glial cells from the intestinal-submucosa using sequential EDTA incubations to chelate divalent cations and then incubation in non-enzymatic cell recovery solution. Plating the resultant cell suspension on poly-D-lysine and laminin results in a highly enriched culture of submucosal glial cells for functional analysis.
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...
FAQs
Removing epithelium first separates the target connective-tissue compartment from the epithelial layer. This allows collagenase and dispase to act on the lamina propria rather than on a mixed tissue sample. The sequence helps enrich the recovered suspension for resident immune, stromal, and other tissue-associated cells, making downstream measurements more interpretable in mucosal studies.
Collagenase and dispase loosen the tissue by breaking down extracellular-matrix proteins and weakening cell-to-matrix and cell-to-cell contacts. This softening allows cells embedded in the connective-tissue layer to separate into a suspension without relying solely on physical disruption. When the treatment preserves viability, the recovered cells remain suitable for analytical and culture-based studies.
It enables researchers to examine resident immune, stromal, and other tissue-associated cells as separate cellular populations rather than only as part of intact tissue. Flow cytometry can support cell analysis, while primary cell culture, microscopy, and single-cell studies provide complementary ways to investigate mucosal biology.
The workflow starts with removal of the epithelium, followed by enzymatic treatment of the underlying connective-tissue layer. Collagenase and dispase then loosen extracellular-matrix and cellular contacts, producing a cell suspension. That suspension can be directed to flow cytometry, primary culture, microscopy, or single-cell analysis, depending on the research question.
The choice of downstream method determines how released cells are characterized. Flow cytometry supports analysis of cellular populations, primary cell culture allows recovered cells to be maintained for further study, microscopy examines their tissue-related context, and single-cell approaches resolve information at the individual-cell level. Together, these options connect cell recovery with distinct experimental readouts.
In medicine and biomedical research, the method provides access to mucosal cells involved in inflammation, immune-cell organization, infection, and tissue responses. Comparing samples from health and disease can help characterize how these cellular populations or their organization relate to altered mucosal biology. Its value lies in linking tissue processing with cellular-level investigation.