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Published on: July 13, 2016
Lineage tracing in the intestinal epithelium
1Hubrecht Institute for Developmental Biology and Stem Cell Research, and University Medical Center Utrecht (UMCU), Utrecht, The Netherlands.
Current Protocols in Stem Cell Biology
|May 6, 2010
Summary
This study details in vivo lineage tracing from Lgr5-positive intestinal stem cells using a specialized mouse model. Protocols allow tracking stem cell contribution to tissue renewal throughout a mouse's lifespan.
Area of Science:
- Stem cell biology
- Developmental biology
- Gastroenterology
Background:
- Lgr5 marks long-lived adult stem cells in various tissues.
- Understanding stem cell contribution to tissue homeostasis is crucial.
- In vivo lineage tracing is essential for studying stem cell dynamics.
Purpose of the Study:
- To describe detailed protocols for in vivo lineage tracing from Lgr5(+) stem cells.
- To utilize the Lgr5-EGFP-ires-CreERT2/Rosa26lacZ mouse model for lineage tracing.
- To demonstrate the application of these protocols in intestinal stem cells and other Lgr5(+) populations.
Main Methods:
- Utilizing an Lgr5-EGFP-ires-CreERT2/Rosa26lacZ mouse model.
- Administering tamoxifen to activate lacZ reporter gene in Lgr5(+) stem cells.
- Tracking the lacZ genetic mark in progeny during homeostasis and tissue renewal.
Main Results:
- Successful initiation of lineage tracing at any age using tamoxifen.
- Permanent genetic marking of Lgr5(+) stem cells and their progeny.
- Demonstration of continuous contribution of Lgr5(+) stem cells to tissue renewal.
Conclusions:
- The described protocols enable robust in vivo lineage tracing from Lgr5(+) stem cells.
- This method allows long-term tracking of stem cell progeny during tissue regeneration.
- The protocols are applicable to intestinal, hair follicle, and stomach stem cell populations.
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