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Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts
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Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts

Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts

DOI:
10.3791/62156-v

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06:57 min

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February 06, 2021

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, , , ,
1Department of Pathology, School of Medicine,University of Michigan

Chapters

  • 00:04Introduction
  • 00:51Preparation of Plates, Chamber Slides and Cell Culture Membrane Inserts
  • 01:30Crypt Isolation
  • 04:09Culturing 2D Monolayer
  • 05:17Results: Reliability of The Primary Epithelial Colon Monolayer Cultures
  • 06:17Conclusion

Summary

Automatic Translation

Automatic Translation

In this protocol, we describe how to generate murine primary epithelial colon monolayers directly from intestinal crypts. We provide experimental approaches to generate confluent monolayers on permeable filters, confluent monolayers for scratch wound healing and biochemical studies, and sparse and confluent monolayers for immunofluorescence analysis.

Tags

Keywords: Primary Colon Epithelial MonolayersIntestinal CryptsWound RepairPermeability BarrierTransepithelial MigrationHost-pathogen InteractionDamaged EpithelialDrug DiscoveryColon DissectionCell IsolationEDTACell Culture

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