The Ly-6A (Sca-1) GFP transgene is expressed in all adult mouse hematopoietic stem cells
The Sca-1 cell surface glycoprotein is used routinely as a marker of adult hematopoietic stem cells (HSCs), allowing a >100-fold enrichment of these rare cells from the bone marrow of the adult mouse. The Sca-1 protein is encoded by the Ly-6A/E gene, a small 4-exon gene that is tightly controlled in its expression in HSCs and several hematopoietic cell types. For the ability to sort and localize HSCs directly from the mouse, we initiated a transgenic approach in which we created Ly-6A (Sca-1) green fluorescent protein (GFP) transgenic mice. We show here that a 14-kb Ly-6A expression cassette directs the transcription of the GFP marker gene in all functional repopulating HSCs in the adult bone marrow. A >100-fold enrichment of HSCs occurred by sorting for the GFP-expressing cells. Furthermore, as shown by fluorescence-activated cell sorting and histologic analysis of several hematopoietic tissues, the GFP transgene expression pattern generally corresponded to that of Sca-1. Thus, the Ly-6A GFP transgene facilitates the enrichment of HSCs and presents the likelihood of identifying HSCs in situ.
|Age Factors, Animals, Antigens, Ly/*genetics, Biological Markers, Bone Marrow Transplantation, Cell Lineage/physiology, Cell Separation/*methods, Gene Expression, Green Fluorescent Proteins, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells/*cytology/*physiology, Indicators and Reagents/metabolism, Luminescent Proteins/genetics, Membrane Proteins/*genetics, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S., Transgenes/genetics|
|Stem Cells: the international journal of cell differentiation and proliferation|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Ma, X, Robin, C.I, Ottersbach, K, & Dzierzak, E.A. (2002). The Ly-6A (Sca-1) GFP transgene is expressed in all adult mouse hematopoietic stem cells. Stem Cells: the international journal of cell differentiation and proliferation. Retrieved from http://hdl.handle.net/1765/10025