X chromosome inactivation in human parthenogenetic embryonic stem cells following prolonged passaging

Qi,Quan; Ding,Chenhui; Hong,Pingping; Yang,Gang; Xie,Yanxin; Wang,Jing; Huang,Sunxing; He,Ke; Zhou,Canquan

doi:10.3892/ijmm.2014.2044

X chromosome inactivation in human parthenogenetic embryonic stem cells following prolonged passaging

Authors:
- Quan Qi
- Chenhui Ding
- Pingping Hong
- Gang Yang
- Yanxin Xie
- Jing Wang
- Sunxing Huang
- Ke He
- Canquan Zhou
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Affiliations: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China
Published online on: December 18, 2014 https://doi.org/10.3892/ijmm.2014.2044
Pages: 569-578
Copyright: © Qi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

The present study aimed to investigate the X chromosome inactivation (XCI) status in long-term cultured human parthenogenetic embryonic stem cells. One human embryonic stem (hES) cell line and 2 human parthenogenetic embryonic stem (hPES) cell lines were subjected to long-term culture in vitro (>50 passages). Karyotyping, array-based comparative genomic hybridization (aCGH), X-inactive specific transcript (XIST) RNA, immunofluorescence staining and real-time PCR were used to assess the chromosome karyotypes of these cells and the XCI status. X chromosome microdeletion was observed in the hPES-2 cells following culture for 50 passages. As early as 20 passages, XIST RNA expression was detected in the hPES-2 cells and was followed by low X-linked gene expression. The XIST RNA expression level was higher in the differentiated hPES-2 cells. The hPES-2' cells that were subclones of hPES-2 retained the XCI status, and had low XIST and X-linked gene expression. XIST RNA expression remained at a low level in the differentiated hPES-2' cells. The human biparental embryonic stem (hBES)-1 and hPES-1 cells did not exhibit XCI, and the differentiated hPES-1 cells had high expression levels of XIST RNA. In conclusion, the chromosome karyotypes of some hPES cell lines revealed instabilities. Similar to the hES cells, the hPES cells exhibited 3 XCI statuses. The unstable XCI status of the hPES-2 line may have been related to chromosome instability. These unstable chromosomes renedered these cells susceptible to environmental conditions and freezing processes, which may be the result of environmental adaptations.

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