Y染色体基因对人类胚胎干细胞自发分化为胚状体的贡献
The Contribution of Y Chromosome Genes to Spontaneous Differentiation of Human Embryonic Stem Cells into Embryoid Bodies .
作者信息
Nafian Dehkordi Simin, Khani Farzaneh, Hassani Seyedeh Nafiseh, Baharvand Hossein, Soleimanpour-Lichaei Hamid Reza, Salekdeh Ghasem Hosseini
机构信息
Department of Stem Cells and Regenerative Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Department of Molecular Systems Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
出版信息
Cell J. 2021 Apr;23(1):40-50. doi: 10.22074/cellj.2021.7145. Epub 2021 Mar 1.
OBJECTIVE
Sexual dimorphism in mammals can be described as subsequent transcriptional differences from their distinct sex chromosome complements. Following X inactivation in females, the Y chromosome is the major genetic difference between sexes. In this study, we used a male embryonic stem cell line (Royan H6) to identify the potential role of the male-specific region of the Y chromosome (MSY) during spontaneous differentiation into embryoid bodies (EBs) as a model of early embryonic development.
MATERIALS AND METHODS
In this experimental study, RH6 cells were cultured on inactivated feeder layers and Matrigel. In a dynamic suspension system, aggregates were generated in the same size and were spontaneously differentiated into EBs. During differentiation, expression patterns of specific markers for three germ layers were compared with MSY genes.
RESULTS
Spontaneous differentiation was determined by downregulation of pluripotent markers and upregulation of fourteen differentiation markers. Upregulation of the ectoderm markers was observed on days 4 and 16, whereas mesoderm markers were upregulated on the 8th day and endodermic markers on days 12-16. Mesoderm markers correlated with 8 MSY genes namely and , which were classified as a mesoderm cluster. Endoderm markers were co-expressed with 7 MSY genes, i.e. and which were grouped as an endoderm cluster. Finally, the ectoderm markers correlated with and genes of the MSY, which were categorized as an ectoderm cluster. In contrast, 2 MSY genes, and were more highly expressed in RH6 cells compared to EBs.
CONCLUSION
We found a significant correlation between spontaneous differentiation and upregulation of specific MSY genes. The expression alterations of MSY genes implied the potential responsibility of their gene co-expression clusters for EB differentiation. We suggest that these genes may play important roles in early embryonic development.
目的
哺乳动物的性二态性可描述为其不同性染色体组成导致的后续转录差异。在雌性个体中X染色体失活后，Y染色体是两性之间主要的遗传差异。在本研究中，我们使用雄性胚胎干细胞系（Royan H6）来确定Y染色体雄性特异性区域（MSY）在自发分化形成胚状体（EBs）过程中的潜在作用，将其作为早期胚胎发育的模型。
材料与方法
在本实验研究中，RH6细胞在灭活的饲养层和基质胶上培养。在动态悬浮系统中，生成大小相同的聚集体并使其自发分化为EBs。在分化过程中，将三个胚层的特异性标志物的表达模式与MSY基因进行比较。
结果
通过多能性标志物的下调和14种分化标志物的上调确定了自发分化。在第4天和第16天观察到外胚层标志物上调，而中胚层标志物在第8天上调，内胚层标志物在第12 - 16天上调。中胚层标志物与8个MSY基因相关，即 和 ，它们被归类为中胚层簇。内胚层标志物与7个MSY基因共表达，即 和 ，它们被归为内胚层簇。最后，外胚层标志物与MSY的 和 基因相关，它们被归类为外胚层簇。相比之下，与EBs相比，2个MSY基因 和 在RH6细胞中表达更高。
结论
我们发现自发分化与特定MSY基因的上调之间存在显著相关性。MSY基因的表达改变暗示其基因共表达簇对EB分化具有潜在作用。我们认为这些基因可能在早期胚胎发育中发挥重要作用。
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