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.
Ma
Xiaoqian
X
Pathology Department, Erasmus University, Rotterdam, Netherlands.
Robin
Catherine
C
Ottersbach
Katrin
K
Dzierzak
Elaine
E
D000367:Age Factors; D000818:Animals; D000950:Antigens, Ly / Q000235:genetics*; D015415:Biomarkers; D016026:Bone Marrow Transplantation; D019070:Cell Lineage / Q000502:physiology; D002469:Cell Separation / Q000379:methods*; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology*; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D019076:Transgenes / Q000235:genetics
D000950:Antigens, Ly; D015415:Biomarkers; D007202:Indicators and Reagents; D008164:Luminescent Proteins; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins