Transcriptomic features of trophoblast lineage cells derived from human induced pluripotent stem cells treated with BMP 4
Transcriptomic features of trophoblast lineage cells derived from human induced pluripotent stem cells treated with BMP 4
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DOI：
10.1016/j.placenta.2019.10.006
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发表时间：
2020-01-01
期刊：
影响因子：
3.8
通讯作者：
Akutsu, Hidenori
中科院分区：
文献类型：
--
作者：
Tsuchida, Nanae;Kojima, Junya;Akutsu, Hidenori
引言：由于缺乏适当的模型系统，对人类胎盘的早期发育仍然知之甚少。以前的报道已经证明，经骨形态发生蛋白4（BMP 4）处理的人诱导多能干细胞（hiPSCs）可以分化为胚外组织，作为滋养层（TB）分化的早期阶段的有用模型。在我们之前的研究中，我们优化了hiPSC衍生的TB谱系的培养条件，但分化的细胞是heterogeneous.Methods：为了表征hiPSC衍生的TB谱系细胞，用50 ng/mL的BMP 4处理四种类型的hiPSC 10天。随后，泛结核标记角蛋白7（KRT 7）阳性的细胞被纯化的分化细胞，使用流式细胞仪和鉴定与DNA microarray.Results：我们的微阵列数据与人类转录组在以前的大规模分析的比较表明，KRT 7+细胞的基因表达模式与胎盘相似。总共有259个上调的基因在所有四个KRT 7+组中普遍表达，包括众所周知的TB标志物。在这些上调的基因中，有几个研究不足的表达模式和功能被证实在原发性胎盘中表达。结论：BMP 4处理的KRT 7+细胞参与了人胎盘的发育过程。此外，这种方法允许鉴定可能参与胎盘形成的新基因。然而，需要进一步的研究来证实它们的功能。
Introduction: Early development of the human placenta remains poorly understood due to the lack of proper model systems. Previous reports have demonstrated that human induced pluripotent stem cells (hiPSCs) treated with bone morphogenetic protein 4 (BMP4) can differentiate into extraembryonic tissues as useful models of the early stage of trophoblast (TB) differentiation. In our previous study, we optimized the culture conditions of hiPSC-derived TB lineages, but the differentiated cells were heterogeneous.Methods: In order to characterize the hiPSC-derived TB lineage cells, four types of hiPSCs were treated with 50 ng/mL of BMP4 for 10 days. Subsequently, cells that were positive for the pan-TB marker keratin 7(KRT7) were purified from the differentiated cells using flow cytometry and identified with a DNA microarray.Results: Comparisons of our microarray data with the human transcriptome in a previous large-scale analysis showed that the gene expression patterns of KRT7+ cells were similar to the placenta. In total, 259 upregulated genes were commonly expressed in all four KRT7+ groups, including well-known TB markers. Among these upregulated genes, several with poorly investigated expression patterns and functions were confirmed as expressed in the primary placenta. While only XAGE2 and KCNQ2 were expressed in TB layers, XAGE2 was expressed throughout pregnancy and KCNQ2 was expressed only in cytotrophoblasts of the first trimester placenta.Conclusion: BMP4-treated KRT7+ cells were in the course of the human placental development. In addition, this approach allowed the identification of new genes that might be involved in placentation. However, further studies are needed to confirm their functions.