Tumor progression of culture-adapted human embryonic stem cells during long-term culture.
Human embryonic stem cells (hESCs) during long-term culture acquire chromosomal changes similar to those occurring in tumorigenesis. This was raised concerns about the progression from hESCs to malignant cells. This study aimed to investigate the changes in chromosomes, cell phenotype, and genes in culture-adapted hESCs to ascertain whether tumorigenic transformation occurred. By cytogenetic analysis we found progressive karyotypic changes from simple to complex in chHES-3, one of the hESC lines established in our laboratory, during a long-term suboptimal culture. We further compared chHES-3 cells at different karyotypic stages in cell surface markers, in vivo differentiation, cell cycle, apoptosis, and gene expression profiles. We found that the karyotypically aberrant chHES-3 had higher S-phase fraction in cell cycle distributions and antiapoptosis ability. In vivo differentiation of karyotypically normal chHES-3 resulted in relatively mature teratoma, whereas karyotypically aberrant chHES-3 formed immature teratoma (grade III), in which more primary neural epithelium was revealed by pathological analysis. The microarray analysis and real-time PCR results showed that some oncogenes were upregulated in karyotypically aberrant chHES-3 cells, whereas the genes related to differentiation were downregulated, and that Wnt signal pathway was activated. In conclusion, chHES-3 cells underwent deregulation of self-renewal and dysfunction of related genes in long-term culture adaptation, leading to malignant transformation.
Yang
Sheng
S
National Center of Human Stem Cell Research and Engineering, Institute of Human Reproduction & Stem Cell Engineering, Central South University, Changsha, China.
Lin
Ge
G
Tan
Yue-Qiu
YQ
Zhou
Di
D
Deng
Lei-Yu
LY
Cheng
De-Hua
DH
Luo
Shu-Wei
SW
Liu
Tian-Cheng
TC
Zhou
Xiao-Ying
XY
Sun
Zheng
Z
Xiang
Yang
Y
Chen
Tian-Ji
TJ
Wen
Ji-Fang
JF
Lu
Guang-Xiu
GX
D018929:Cell Culture Techniques; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology; D002478:Cells, Cultured; D020732:Cytogenetic Analysis; D053595:Embryonic Stem Cells / Q000473:pathology*; D020869:Gene Expression Profiling; D006801:Humans; D009385:Neoplastic Processes; D016196:S Phase / Q000235:genetics