Cell line-dependent differentiation of induced pluripotent stem cells into cardiomyocytes in mice
Cell line-dependent differentiation of induced pluripotent stem cells into cardiomyocytes in mice
复制标题
DOI：
10.1093/cvr/cvq189
复制
发表时间：
2010-11-01
影响因子：
10.8
通讯作者：
Heike, Toshio
中科院分区：
文献类型：
--
作者：
Kaichi, Shinji;Hasegawa, Koji;Heike, Toshio
小鼠和人成纤维细胞可以通过四种转录因子（Oct 3/4、Sox 2、Klf 4和c-Myc）的异位表达直接重编程为多能性，以产生诱导的多能干（iPS）细胞。即使不表达c-Myc也可以产生iPS细胞。本研究在三种不同的细胞系中检测了小鼠iPS细胞向心肌细胞的分化模式，在无饲养层的明胶化培养皿上诱导分化过程中，与心脏发生相关的基因表达与胚胎干细胞相同，并且在两种iPS细胞系中发生了肌源性收缩。然而，在一个iPS细胞系（20 D17）由四个因素，心脏特异性基因的表达和搏动活动是非常低的。用组蛋白去乙酰化酶（HDAC）抑制剂-虽然基础Nkx2.5表达在20 D17非常低，但TSA诱导的增加最大。TSA在20 D17也诱导了收缩蛋白的表达。此外，我们还证实，与其他两种iPS细胞系相比，20 D17中Oct 3/4的mRNA水平和HDAC 4的核蛋白水平增加。DNA微阵列分析鉴定了20 D17表达上调或下调的基因。小鼠iPS细胞以细胞系依赖的方式分化为心肌细胞。TSA诱导小鼠iPS细胞的心肌分化，可能有助于克服分化效率的细胞系差异。
Mouse and human fibroblasts can be directly reprogrammed to pluripotency by the ectopic expression of four transcription factors (Oct3/4, Sox2, Klf4, and c-Myc) to yield induced pluripotent stem (iPS) cells. iPS cells can be generated even without the expression of c-Myc. The present study examined patterns of differentiation of mouse iPS cells into cardiomyocytes in three different cell lines reprogrammed by three or four factors.During the induction of differentiation on feeder-free gelatinized dishes, genes involved in cardiogenesis were expressed as in embryonic stem cells and myogenic contraction occurred in two iPS cell lines. However, in one iPS cell line (20D17) generated by four factors, the expression of cardiac-specific genes and the beating activity were extremely low. Treating iPS cells with trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, increased Nkx2.5 expression in all iPS cell lines. While the basal Nkx2.5 expression was very low in 20D17, the TSA-induced increase was the greatest. TSA also induced the expression of contractile proteins in 20D17. Furthermore, we demonstrated the increased mRNA level of Oct3/4 and nuclear protein level of HDAC4 in 20D17 compared with the other two iPS cell lines. DNA microarray analysis identified genes whose expression is up- or down-regulated in 20D17.Mouse iPS cells differentiate into cardiomyocytes in a cell line-dependent manner. TSA induces myocardial differentiation in mouse iPS cells and might be useful to overcome cell line variation in the differentiation efficiency.