title: X chromosome inactivation in human parthenogenetic embryonic stem cells following prolonged passaging.
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.
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Qi
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Chenhui
authors.initials: C
authors.lastname: Ding
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Hong
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Gang
authors.initials: G
authors.lastname: Yang
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Yanxin
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Sunxing
authors.initials: S
authors.lastname: Huang
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Ke
authors.initials: K
authors.lastname: He
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China.
authors.forename: Canquan
authors.initials: C
authors.lastname: Zhou
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D050172:Genes, X-Linked; D042822:Genomic Instability; D006657:Histones / Q000378:metabolism; D006801:Humans; D059785:Karyotype; D010312:Parthenogenesis / Q000235:genetics*; D062085:RNA, Long Noncoding / Q000235:genetics; D049951:X Chromosome Inactivation
chemical_list: D006657:Histones; D062085:RNA, Long Noncoding; C077885:XIST non-coding RNA