Immunophenotyping of Live Human Pluripotent Stem Cells by Flow Cytometry
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Human pluripotent stem cells (hPSCs) have great potential for use in regenerative medicine and cell replacement therapies; however, prior to clinical application, cultured cell populations need to be screened to ensure the quality of the culture, as well as the capacity of these pluripotent cells to differentiate into desired cell types. Flow cytometry, utilizing antibodies recognizing targets restricted to the hPSC surfaceome, offers an invaluable tool for high-throughput validation of hPSC lines. Here we describe the immunophenotyping of live human embryonic stem cell (hESC, H9) and human induced pluripotent stem cell (hiPSC, KB3) lines by flow cytometry using a panel of antibodies identified as either stem cell reference markers (CD90, EpCam) or reported as being prevalent or restricted (c-Kit, HPI-1, Integrin α6, Semaphorin-6A) to these cells. The protocols described here with hPSCs are also applicable to differentiated hPSC progeny and should be instrumental in the immunophenotyping and isolation of well-defined homogeneous cell populations useful in regenerative medicine.
Key wordsEmbryonic stem cells Induced pluripotent stem cells Surfaceome Immunophenotyping Flow cytometry Antibody
This research was supported by the Intramural Research Program of the NIH, National Institute on Aging, by the Research Grants Council of Hong Kong Theme-based Research Scheme T13-706/11, and the Hong Kong Research Grant Committee General Research Fund (Project number 17100214). We thank Robert Wersto and the NIA FC Core Facility for assistance with flow cytometry.
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