|Title||The Ly-6A (Sca-1) GFP transgene is expressed in all adult mouse hematopoietic stem cells.|
|Publication Type||Journal Article|
|Year of Publication||2002|
|Authors||Ma X, Robin C, Ottersbach K, Dzierzak E|
|Keywords||Age Factors, Animals, Antigens, Ly, Biological Markers, Bone Marrow Transplantation, Cell Lineage, Cell Separation, Gene Expression, Green Fluorescent Proteins, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Indicators and Reagents, Luminescent Proteins, Membrane Proteins, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, Transgenes|
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.
|Alternate Journal||Stem Cells|
|Grant List||DK51077 / DK / NIDDK NIH HHS / United States|