A Novel Strategy for Isolation, Molecular and Functional Characterization of Embryonic Mammary Stem Cells Using Molecular Genetics and Microfluidic Sorting
Abstract
We developed a genetic system to identify, isolate, and characterize mammary stem cells. Our system consists of an activator component that is dependent on Wnt signaling, which is essential for mammary gland development, and on a doxycycline regulated reporter to toggle the system on and off. The reporter component labels the chromatin of cells for direct visualization. We used a modular design to enable the system to be applied to cancer models and other organs. Over the past year we have targeted mouse embryonic stem cells with our modular transgenic system and validated its Wnt responsiveness. We have also established assays for isolation and characterization of small numbers of viable, fluorescently labeled cells. In parallel, we have determined that embryonic mammary rudiments contain concentrated mammary stem cells and are isolating these structures to obtain stem cell specific expression signatures. 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, 2008
- Accession Number
- ADA490322
Entities
People
- Geoffrey M Wahl
Organizations
- Salk Institute for Biological Studies