Low microRNA-199a expression in human amniotic epithelial cell feeder layers maintains human-induced pluripotent stem cell pluripotency via increased leukemia inhibitory factor expression
Low microRNA-199a expression in human amniotic epithelial cell feeder layers maintains human-induced pluripotent stem cell pluripotency via increased leukemia inhibitory factor expression
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DOI：
10.1093/abbs/gmr127
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发表时间：
2012-03-01
影响因子：
3.7
通讯作者：
Guo, Lihe
中科院分区：
文献类型：
--
作者：
Liu, Te;Chen, Qing;Guo, Lihe
人类诱导的多能干细胞（iPS）与胚胎干细胞具有相同的关键特性，并且可以从患者或疾病特异性来源产生，这使得它们对个性化医疗，药物筛选或细胞治疗具有吸引力。长期培养和维持正常iPS细胞处于未分化的自我更新状态是一个重大挑战。我们之前的研究表明，人羊膜上皮细胞（waecs）可以为小鼠和人胚胎干细胞或精原干细胞提供良好的饲养细胞来源，因为它们高水平表达内源性白血病抑制因子（LIF）。在这里，我们研究了外源性microRNA-199a调控对huaec内源性LIF表达的影响，进而对人iPS细胞多能性的影响。我们发现转染microRNA-199a突变体的huaec饲养细胞高水平表达LIF，使iPS在长期培养中保持高水平的碱性磷酸酶活性，并在严重联合免疫缺陷小鼠中形成畸胎瘤。转染microRNA-199a突变体的诱导多能干细胞与转染microRNA-199a的诱导多能干细胞或小鼠胚胎成纤维细胞的诱导多能干细胞的表达明显增加。综上所述，这些结果表明，LIF的表达可能受microRNA-199a的调控，LIF是饲养细胞的关键成分，也是维持人类iPS细胞处于未分化、增殖、自我更新状态所必需的。
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.