A Novel Strategy for Isolation, Molecular and Functional Characterization of Embryonic Mammary Stem Cells Using Molecular Genetics and Microfluidic Sorting
Abstract
We are developing a genetic system to identify isolate and characterize mammary stem cells. Our system consists of both activator and reporter components. The activator component is dependent both on Wnt signaling which is essential for mammary gland development and on doxycycline to toggle the system on and off. The reporter component labels cells for direct visualization. We used a modular design to enable the system to be applied to cancer models and other organs. Data obtained over the past year with a related proof of concept system showed that mammary glands were brightly labeled by H2BGFP fluorescence in response to the Tet transactivator. This initial evaluation demonstrates the feasibility of the approach in vivo. In vitro analyses showed that chromatin is labeled by H2BGFP regardless of cell cycle phase. The molecular reagents and strategies we are developing have broad applications for studies examining the relationship between normal and cancer stem cells and determining whether they share the same origin.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 2007
- Accession Number
- ADA474943
Entities
People
- Geoffrey M Wahl
Organizations
- Salk Institute for Biological Studies