Department of Regenerative Medicine and Advanced Cardiac Therapeutics, Keio University School of Medicine, Tokyo, Japan.
Nat Methods. 2010 Jan;7(1):61-6. doi: 10.1038/nmeth.1403. Epub 2009 Nov 29.
Several applications of pluripotent stem cell (PSC)-derived cardiomyocytes require elimination of undifferentiated cells. A major limitation for cardiomyocyte purification is the lack of easy and specific cell marking techniques. We found that a fluorescent dye that labels mitochondria, tetramethylrhodamine methyl ester perchlorate, could be used to selectively mark embryonic and neonatal rat cardiomyocytes, as well as mouse, marmoset and human PSC-derived cardiomyocytes, and that the cells could subsequently be enriched (>99% purity) by fluorescence-activated cell sorting. Purified cardiomyocytes transplanted into testes did not induce teratoma formation. Moreover, aggregate formation of PSC-derived cardiomyocytes through homophilic cell-cell adhesion improved their survival in the immunodeficient mouse heart. Our approaches will aid in the future success of using PSC-derived cardiomyocytes for basic and clinical applications.
多能干细胞（PSC）衍生的心肌细胞的几种应用需要消除未分化的细胞。心肌细胞纯化的一个主要限制是缺乏简单和特异的细胞标记技术。我们发现，一种标记线粒体的荧光染料，四甲基罗丹明甲酯高氯酸盐，可以用于选择性标记胚胎和新生大鼠心肌细胞，以及小鼠、狨猴和人 PSC 衍生的心肌细胞，并且可以通过荧光激活细胞分选将细胞进行富集（纯度>99%）。移植到睾丸中的纯化心肌细胞不会诱导畸胎瘤形成。此外，PSC 衍生的心肌细胞通过同源细胞-细胞黏附形成聚集体，提高了它们在免疫缺陷小鼠心脏中的存活率。我们的方法将有助于未来使用 PSC 衍生的心肌细胞进行基础和临床应用的成功。
