SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells.
To identify cell-surface markers specific to human cardiomyocytes, we screened cardiovascular cell populations derived from human embryonic stem cells (hESCs) against a panel of 370 known CD antibodies. This screen identified the signal-regulatory protein alpha (SIRPA) as a marker expressed specifically on cardiomyocytes derived from hESCs and human induced pluripotent stem cells (hiPSCs), and PECAM, THY1, PDGFRB and ITGA1 as markers of the nonmyocyte population. Cell sorting with an antibody against SIRPA allowed for the enrichment of cardiac precursors and cardiomyocytes from hESC/hiPSC differentiation cultures, yielding populations of up to 98% cardiac troponin T-positive cells. When plated in culture, SIRPA-positive cells were contracting and could be maintained over extended periods of time. These findings provide a simple method for isolating populations of cardiomyocytes from human pluripotent stem cell cultures, and thereby establish a readily adaptable technology for generating large numbers of enriched cardiomyocytes for therapeutic applications.
McEwen Centre for Regenerative Medicine, Ontario Cancer Institute, University Health Network, Toronto, Ontario, Canada.
Nicole C
NC
Dubois
April M
AM
Craft
Parveen
P
Sharma
David A
DA
Elliott
Edouard G
EG
Stanley
Andrew G
AG
Elefanty
Anthony
A
Gramolini
Gordon
G
Keller
D000943:Antigens, Differentiation / Q000378:metabolism*; D000954:Antigens, Surface / Q000032:analysis; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism*
D000943:Antigens, Differentiation; D000954:Antigens, Surface; D015415:Biomarkers; D011971:Receptors, Immunologic; C501920:SIRPA protein, human