Riordon Daniel R, Boheler Kenneth R
Laboratory of Cardiovascular Sciences, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA.
School of Biomedical Sciences, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, SAR, China.
Methods Mol Biol. 2018;1722:127-149. doi: 10.1007/978-1-4939-7553-2_9.
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.
人类多能干细胞（hPSC）在再生医学和细胞替代疗法中具有巨大的应用潜力；然而，在临床应用之前，需要对培养的细胞群体进行筛选，以确保培养物的质量以及这些多能细胞分化为所需细胞类型的能力。利用识别仅限于hPSC表面组靶标的抗体进行流式细胞术，为hPSC系的高通量验证提供了一种非常有价值的工具。在这里，我们描述了通过流式细胞术对活的人类胚胎干细胞（hESC，H9）和人类诱导多能干细胞（hiPSC，KB3）系进行免疫表型分析，使用一组被鉴定为干细胞参考标志物（CD90、EpCam）或据报道在这些细胞中普遍存在或受限的抗体（c-Kit、HPI-1、整合素α6、信号素-6A）。这里描述的hPSC实验方案也适用于分化的hPSC后代，并且在再生医学中用于明确的同质细胞群体的免疫表型分析和分离方面应该很有帮助。
