Low microRNA-199a expression in human amniotic epithelial cell feeder layers maintains human-induced pluripotent stem cell pluripotency via increased leukemia inhibitory factor expression

• Published in: Acta biochimica et biophysica Sinica Vol. 44; no. 3; pp. 197 - 206
• Main Authors: Liu, Te, Chen, Qing, Huang, Yongyi, Huang, Qin, Jiang, Lizhen, Guo, Lihe
• Format: Journal Article
• Language: English
• Published: China 01.03.2012
• Subjects: Amniotic Fluid - cytology; Animals; Cell Culture Techniques - methods; Coculture Techniques - methods; Culture Techniques - methods; Epithelial Cells - cytology; Fibroblasts - cytology; Gene Expression Regulation; Humans; Leukemia Inhibitory Factor - biosynthesis; Leukemia Inhibitory Factor - metabolism; Mice; Mice, Inbred C57BL; MicroRNAs - biosynthesis; MicroRNAs - physiology; Pluripotent Stem Cells - cytology; Stem Cells - cytology; 上皮细胞; 人类胚胎干细胞; 多能干细胞; 滋养层细胞; 白血病抑制因子; 维护; 羊膜; 诱导
• ISSN: 1672-9145; 1745-7270
• DOI: 10.1093/abbs/gmr127

Human-induced pluripotent stem （iPS） cells share the same key properties as embryonic stem cells, and may be gener- ated from patient- or disease-specific sources, which makes them attractive for personalized medicine, drug screens, or cellular therapy. Long-term cultivation and maintenance of normal iPS cells in an undifferentiated self-renewing state is a major challenge. Our previous studies have shown that human amniotic epithelial cells 0IuAECs） could provide a good source of feeder cells for mouse and human embryonic stem cells, or spermatogonial stem cells, as they express endogenous leukemia inhibitory factor （LIF） at high levels. Here, we examined the effect of exogenous microRNA-199a regulation on endogenous LIF expression in HuAECs, and in turn on human iPS cell pluripotency. We found that HuAECs feeder cells transfected with microRNA-199a mutant expressed LIF at high levels, allowing iPS to main- tain a high level of alkaline phosphatase activity in long- term culture and form teratomas in severe combined immunodeficient mice. The expression of stem cell markers was increased in iPS cultured on HuAECs feeder cells trans- fected with the microRNA-199a mutant, compared with iPS cultured on HuAECs transfected with microRNA-199a or mouse embryo fibroblasts. Taken together, these results suggested that LIF expression might be regulated by microRNA-199a, and LIF was a crucial component in feeder cells, and also was required for maintenance of human iPS cells in an undifferentiated, proliferative state capable of serf-renewaL