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The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells
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Chapters
- 00:05Title
- 01:10Isolation of Small Intestinal Crypts
- 04:02Single Cell Preparation, Flow Cytometry, and Plating
- 07:34Results: Effect of siRNA Medicated Knockdown of Apc in Lgr5+ Cells
- 08:29Conclusion
Intestinal organoid cultures are established from whole crypts and do not allow the analysis of self-renewal and differentiation in a cell-specific fashion. This protocol describes reconstitution of sorted stem (Lgr5+) and niche (Paneth) cells, which give rise to organoids while enabling their prior biochemical and genetic modification and functional analysis.
