Direct differentiation of hepatic cells from human induced pluripotent stem cells using a limited number of cytokines
Direct differentiation of hepatic cells from human induced pluripotent stem cells using a limited number of cytokines
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DOI：
10.1007/s12072-011-9251-5
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发表时间：
2011-12-01
影响因子：
6.6
通讯作者：
Koike, Kazuhiko
中科院分区：
文献类型：
--
作者：
Takata, Akemi;Otsuka, Motoyuki;Koike, Kazuhiko
目的改进诱导多能干细胞（iPS）分化为肝细胞的方法是其在肝病学领域应用的重要一步。具体而言，应减少不同细胞因子的数量以限制不期望的作用并降低该方法的成本。在这份报告中，我们描述了一个简单的方法，用于指导人类iPS细胞分化成肝细胞，只使用两种细胞因子和一个短的孵育时间。施加高剂量的激活素A 3天以诱导定形内胚层形成。随后，用肝细胞生长因子（HGF）处理细胞5天以产生肝细胞。通过分化标记物的免疫染色确认分化。白蛋白的mRNA水平在分化的肝细胞产生的使用先前测试的三步协议，使用激活素A，成纤维细胞生长因子（FGF）/骨形态发生蛋白（BMP），和肝细胞生长因子，我们的新协议进行了比较，以确定分化的效率。结果我们的两步协议诱导的iPS细胞分化成肝细胞，需要较短的分化期比以前的三步协议。两种方案的分化效率是可比的，诱导的肝细胞functional.Conclusions开发高效的诱导和培养方法，以产生更高度成熟的肝细胞是必不可少的再生细胞为基础的治疗。我们的方案提供了一种简单、经济、省时的方法，用于从iPS细胞产生肝细胞。
Purpose Development of improved protocols for differentiating induced pluripotent stem (iPS) cells into hepatic cells is an important step toward their use in the field of hepatology. Specifically, the number of different cytokines should be reduced to limit undesired effects and to reduce the cost of the process. In this report, we describe a simple method for directing human iPS cells to differentiate into hepatic cells using only two cytokines and a short incubation time.Methods A two-step protocol for differentiating iPS cells into hepatic cells was developed. A high dose of activin A was applied for 3 days to induce definitive endoderm formation. Subsequently, cells were treated with hepatocyte growth factor (HGF) for 5 days to generate hepatic cells. Differentiation was confirmed by immunostaining for differentiation markers. Albumin mRNA levels in differentiated hepatic cells generated using a previously tested three-step protocol that uses activin A, fibroblast growth factor (FGF)/bone morphogenetic protein (BMP), and HGF, and our new protocol were compared to determine the efficiency of differentiation.Results Our two-step protocol induced the differentiation of iPS cells into hepatic cells and required a shorter differentiation period than the previous three-step protocol. The differentiation efficiencies of the two protocols were comparable and the induced hepatic cells were functional.Conclusions Developing efficient induction and culture methods to generate more highly matured hepatocytes is essential for regenerative cell-based therapies. Our protocol provides a simple, cost-effective, and time-saving approach for generating hepatic cells from iPS cells.