McEwen Centre for Regenerative Medicine, Ontario Cancer Institute, University Health Network, Toronto, Ontario, Canada.
Nat Biotechnol. 2011 Oct 23;29(11):1011-8. doi: 10.1038/nbt.2005.
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.
为了鉴定人类心肌细胞特有的细胞表面标志物，我们用一组 370 种已知的 CD 抗体对来源于人类胚胎干细胞（hESC）的心血管细胞群体进行了筛选。该筛选鉴定出信号调节蛋白 α（SIRPA）是一种特异性表达于来源于 hESC 和人诱导多能干细胞（hiPSC）的心肌细胞上的标志物，而 PECAM、THY1、PDGFRB 和 ITGA1 则是心肌细胞以外的细胞群体的标志物。用针对 SIRPA 的抗体进行细胞分选，可从 hESC/hiPSC 分化培养物中富集心脏前体细胞和心肌细胞，得到高达 98%的心肌肌钙蛋白 T 阳性细胞群体。当在培养中铺板时，SIRPA 阳性细胞在收缩，并能在较长时间内维持。这些发现为从人类多能干细胞培养物中分离心肌细胞群体提供了一种简单的方法，并因此建立了一种易于适应的技术，可用于产生大量用于治疗应用的富集心肌细胞。
