Journal
/
Bioengineering
/
The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells
JoVE Journal
Bioengineering
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Bioengineering
The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells

The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells

DOI:
10.3791/56329-v

•

09:38 min

•

November 20, 2017

•

, , ,
1Department of Pathology, Erasmus MC Cancer Institute,Erasmus University Medical Center

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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Intestinal Stem CellsLgr5Paneth CellsOrganoid Reconstitution AssayIntestinal NicheCell IsolationCell SortingIntestinal Crypts

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Use of Human Perivascular Stem Cells for Bone Regeneration

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Transplantation of Induced Pluripotent Stem Cell-derived Mesoangioblast-like Myogenic Progenitors in Mouse Models of Muscle Regeneration

A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells

Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis

Optimizing Attachment of Human Mesenchymal Stem Cells on Poly(ε-caprolactone) Electrospun Yarns

Protocol for MicroRNA Transfer into Adult Bone Marrow-derived Hematopoietic Stem Cells to Enable Cell Engineering Combined with Magnetic Targeting

A Quantitative Fluorescence Microscopy-based Single Liposome Assay for Detecting the Compositional Inhomogeneity Between Individual Liposomes

Bioparticle Microarrays for Chemotactic and Molecular Analysis of Human Neutrophil Swarming in vitro

Isolation, Expansion, and Differentiation of Mesenchymal Stem Cells from the Infrapatellar Fat Pad of the Goat Stifle Joint

Read Article