The dynamic changes of X chromosome inactivation during early culture of human embryonic stem cells.
X chromosome inactivation (XCI) is required for dosage compensation of X-linked genes in human female cells. Several previous reports have described the promiscuous XCI status in long-term cultured female human embryonic stem cells (hESCs), and the majority of them exhibit non-random XCI. However, when and how such female hESCs acquire the aberrant XCI states during culture is unknown. Herein, through comparing the XCI states in 18 paired hES cell lines throughout early culture, we revealed a uniform dynamic change during this culture period under a widely used culture condition. The female initial hESCs (ihESCs, P4-P9) expressed XIST RNA, H3K27me3 punctate enrichment and displayed random XCI pattern. By further culturing, the female early hESCs (ehESCs, P20-P30) lost the expression of XIST RNA, H3K27me3 punctate enrichment and exhibited a completely skewed XCI pattern. Importantly, a subset of X-linked genes was up-regulated in ehESCs, including some cancer-related genes. At last, we found 5% physiological oxygen was beneficial for the expression of XIST and H3K27me3 punctate enrichment, but not for the XCI pattern. We conclude that the XCI dynamic change is a frequent epigenetic instability event during early culture, which is accompanied by the up-regulation of some X-linked genes. Furthermore, we emphasize that physiological oxygen is beneficial for XCI fidelity.
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Pingyuan
P
Xie
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Qi
Q
Ouyang
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Lizhi
L
Leng
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Liang
L
Hu
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Dehua
D
Cheng
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Yueqiu
Y
Tan
National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China.
Guangxiu
G
Lu
Institute of Reproductive & Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha 410078, China; National Engineering and Research Center of Human Stem Cell, Changsha 410078, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha 410078, China. Electronic address: linggf@hotmail.com.
Ge
G
Lin
D002454:Cell Differentiation; D002478:Cells, Cultured; D019175:DNA Methylation; D005260:Female; D006657:Histones / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D010100:Oxygen / Q000378:metabolism; D011401:Promoter Regions, Genetic; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism; D049951:X Chromosome Inactivation
D006657:Histones; D062085:RNA, Long Noncoding; C077885:XIST non-coding RNA; D010100:Oxygen
Culture alteration; Epigenetics; Gene expression regulation; Human embryonic stem cells; X chromosome inactivation