Low microRNA-199a expression in human amniotic epithelial cell feeder layers maintains human-induced pluripotent stem cell pluripotency via increased leukemia inhibitory factor expression
| Autor: | Qing Chen, Te Liu, Yongyi Huang, Lizhen Jiang, Lihe Guo, Qin Huang |
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| Rok vydání: | 2012 |
| Předmět: |
Pluripotent Stem Cells
KOSR Cell Culture Techniques Biophysics Biology Stem cell marker Leukemia Inhibitory Factor Biochemistry Cell therapy Mice Culture Techniques Animals Humans Induced pluripotent stem cell Stem Cells Epithelial Cells General Medicine Fibroblasts Amniotic Fluid Embryonic stem cell Coculture Techniques Cell biology Mice Inbred C57BL MicroRNAs Gene Expression Regulation Amniotic epithelial cells embryonic structures Immunology Stem cell Leukemia inhibitory factor |
| Zdroj: | Acta Biochimica et Biophysica Sinica. 44:197-206 |
| ISSN: | 1672-9145 |
| Popis: | Human-induced pluripotent stem (iPS) cells share the same key properties as embryonic stem cells, and may be generated 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 (HuAECs) 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 maintain 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 transfected 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 self-renewal. |
| Databáze: | OpenAIRE |
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