Gene Therapy and Hepatology Area, FIMA University of Navarra, Pamplona, Spain.
J Physiol Biochem. 2013 Dec;69(4):835-45. doi: 10.1007/s13105-013-0260-9. Epub 2013 May 30.
Hepatocyte transplantation is considered a promising therapy for patients with liver diseases. Induced pluripotent stem cells (iPSCs) are an unlimited source for the generation of functional hepatocytes. While several protocols that direct the differentiation of iPSCs into hepatocyte-like cells have already been reported, the liver engraftment potential of iPSC progeny obtained at each step of hepatic differentiation has not yet been thoroughly investigated. In this study, we present an efficient strategy to differentiate mouse iPSCs into hepatocyte-like cells and evaluate their liver engraftment potential at different time points of the protocol (5, 10, 15, and 20 days of differentiation). iPSCs were differentiated in the presence of cytokines, growth factors, and small molecules to finally generate hepatocyte-like cells. These iPSC-derived hepatocyte-like cells exhibited hepatocyte-associated functions, such as albumin secretion and urea synthesis. When we transplanted iPSC progeny into the spleen, we found that 15- and 20-day iPSC progeny engrafted into the livers and further acquired hepatocyte morphology. In contrast, 5- and 10-day iPSC progeny were also able to engraft but did not generate hepatocyte-like cells in vivo. Our data may aid in improving current protocols geared towards the use of iPSCs as a new source of liver-targeted cell therapies.
肝细胞移植被认为是治疗肝脏疾病患者的一种有前途的疗法。诱导多能干细胞（iPSCs）是产生功能性肝细胞的无限来源。虽然已经报道了几种将 iPSCs 定向分化为肝样细胞的方案，但尚未彻底研究 iPSC 后代在肝分化的每个步骤中的肝定植潜力。在这项研究中，我们提出了一种有效的策略，可将小鼠 iPSCs 分化为肝样细胞，并在方案的不同时间点（分化的第 5、10、15 和 20 天）评估其肝定植潜力。在细胞因子、生长因子和小分子的存在下将 iPSCs 分化，最终生成肝样细胞。这些 iPSC 衍生的肝样细胞表现出与肝细胞相关的功能，如白蛋白分泌和尿素合成。当我们将 iPSC 后代移植到脾脏中时，我们发现 15 天和 20 天的 iPSC 后代定植到肝脏中，并进一步获得了肝细胞形态。相比之下，5 天和 10 天的 iPSC 后代也能够定植，但在体内未产生肝样细胞。我们的数据可能有助于改进目前使用 iPSCs 作为肝脏靶向细胞治疗新来源的方案。
