High Oxygen Condition Facilitates the Differentiation of Mouse and Human Pluripotent Stem Cells into Pancreatic Progenitors and Insulin-producing Cells*
High Oxygen Condition Facilitates the Differentiation of Mouse and Human Pluripotent Stem Cells into Pancreatic Progenitors and Insulin-producing Cells*
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DOI：
10.1074/jbc.m113.524363
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发表时间：
2014-04-04
影响因子：
4.8
通讯作者：
Tomizawa, Kazuhito
中科院分区：
文献类型：
--
作者：
Hakim, Farzana;Kaitsuka, Taku;Tomizawa, Kazuhito
背景：氧气在器官发育中发挥着关键作用，包括胰腺细胞。结果：高氧条件会增加小鼠和人类多能干细胞的 Ngn3 阳性和胰岛素阳性细胞。结论：高氧条件下培养对胰腺分化有促进作用。意义：这项新技术提供了一种利用患者特异性iPS细胞治疗糖尿病的有效方法。多能干细胞在糖尿病再生医学中具有潜在的应用前景。干细胞分化为产生胰岛素的细胞已通过多种方案实现。然而，该方法的效率和分化细胞的效力都不足。氧张力（氧分压）已被证明可以调节包括胰腺细胞在内的多个器官的胚胎发育。在这项研究中，我们试图建立一种通过在高氧（O-2）条件下培养诱导多能干细胞（iPSC）分化为胰岛素产生细胞的有效方法。在分化早期采用高 O-2 条件治疗会增加分化末期的胰岛素阳性细胞。我们发现，在胰腺祖细胞阶段，高 O-2 条件抑制了 Notch 依赖性基因 Hes1 的表达，并增加了 Ngn3 的表达。这种作用是通过抑制缺氧诱导因子 1 蛋白水平引起的。此外，高 O-2 条件会激活 Wnt 信号传导。采用高 O-2 条件进行最佳阶段特异性治疗可显着增加小鼠胚胎干细胞和人 iPSC 中的胰岛素产量，并在人 iPSC 中产生含有高达 10% C 肽阳性细胞的群体。这些结果表明，在特定阶段的高 O-2 条件下培养对于胰岛素生成细胞的有效生成是有用的。
Background: Oxygen plays a key role in organ development, including pancreatic -cells. Results: High oxygen conditions increase Ngn3-positive and insulin-positive cells from both mouse and human pluripotent stem cells. Conclusion: Culturing under high oxygen conditions has a facilitative effect on pancreatic differentiation. Significance: This new technique provides an efficient method to utilize patient-specific iPS cells for the treatment of diabetes.Pluripotent stem cells have potential applications in regenerative medicine for diabetes. Differentiation of stem cells into insulin-producing cells has been achieved using various protocols. However, both the efficiency of the method and potency of differentiated cells are insufficient. Oxygen tension, the partial pressure of oxygen, has been shown to regulate the embryonic development of several organs, including pancreatic -cells. In this study, we tried to establish an effective method for the differentiation of induced pluripotent stem cells (iPSCs) into insulin-producing cells by culturing under high oxygen (O-2) conditions. Treatment with a high O-2 condition in the early stage of differentiation increased insulin-positive cells at the terminus of differentiation. We found that a high O-2 condition repressed Notch-dependent gene Hes1 expression and increased Ngn3 expression at the stage of pancreatic progenitors. This effect was caused by inhibition of hypoxia-inducible factor-1 protein level. Moreover, a high O-2 condition activated Wnt signaling. Optimal stage-specific treatment with a high O-2 condition resulted in a significant increase in insulin production in both mouse embryonic stem cells and human iPSCs and yielded populations containing up to 10% C-peptide-positive cells in human iPSCs. These results suggest that culturing in a high O-2 condition at a specific stage is useful for the efficient generation of insulin-producing cells.