Kimura Maya, Furukawa Hatsue, Shoji Masanobu, Shinozawa Tadahiro
Drug Safety Research Laboratories.
Integrated Technology Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1 Muraoka-Higashi 2-chome, Fujisawa, Kanagawa 251-8555, Japan.
J Stem Cells Regen Med. 2019 Dec 24;15(2):45-51. doi: 10.46582/jsrm.1502009. eCollection 2019.
Human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) have attracted attention as a novel tool for drug safety screening and several differentiation protocols of hiPSC lines into cardiomyocytes have been reported; the standardization of these protocols will expand their applications for safety assessments such as "clinical safety trial-on-dish". Bone morphogenetic protein 4 (BMP4) is an important factor in promoting mesoderm differentiation and BMP4 treatment has been used at the early stage of cardiac differentiation into different hiPSCs. In the present study, we evaluated the effects of BMP4 treatment at the early stage of cardiac differentiation. We performed gene expression profiling of the germ layer during mesoderm differentiation of hiPSCs derived from three different donors. The expression of (a mesoderm marker) and (an endoderm marker) increased and that of (a neuroectoderm marker) decreased in pooled embryoid bodies (EBs) after BMP4 treatment. Single-cell gene expression analysis revealed that mesodermal and mesendodermal populations increased in EBs derived from 253G1. Finally, BMP4 treatment increased mesodermal and mesendodermal populations compared with that without BMP4 in two other hiPSC lines, confirming the reproducibility of multiple hiPSC lines. Thus, our results suggest that BMP4 treatment increases mesodermal and mesendodermal populations at the early stage of cardiac differentiation in different hiPSC lines.
人诱导多能干细胞衍生的心肌细胞（hiPS-CMs）作为药物安全性筛选的新型工具已受到关注，并且已有数种将hiPSC系分化为心肌细胞的方案被报道；这些方案的标准化将扩大其在诸如“盘中临床安全性试验”等安全性评估中的应用。骨形态发生蛋白4（BMP4）是促进中胚层分化的重要因子，在不同hiPSC的心脏分化早期已使用BMP4处理。在本研究中，我们评估了心脏分化早期BMP4处理的效果。我们对来自三个不同供体的hiPSC在中胚层分化过程中的胚层进行了基因表达谱分析。BMP4处理后，汇集的胚状体（EBs）中（中胚层标志物）和（内胚层标志物）的表达增加，而（神经外胚层标志物）的表达降低。单细胞基因表达分析显示，来自253G1的EBs中中胚层和中内胚层群体增加。最后，与另外两个hiPSC系中未使用BMP4处理相比，BMP4处理增加了中胚层和中内胚层群体，证实了多个hiPSC系的可重复性。因此，我们的结果表明，BMP4处理在不同hiPSC系的心脏分化早期增加了中胚层和中内胚层群体。
