Isolation and expansion of human pluripotent stem cell-derived hepatic progenitor cells by growth factor defined serum-free culture conditions
Isolation and expansion of human pluripotent stem cell-derived hepatic progenitor cells by growth factor defined serum-free culture conditions
复制标题
DOI：
10.1016/j.yexcr.2017.02.022
复制
发表时间：
2017-03-15
影响因子：
3.7
通讯作者：
Furue, Miho K.
中科院分区：
文献类型：
--
作者：
Fukuda, Takayuki;Takayama, Kazuo;Furue, Miho K.
有限的生长潜力、狭窄的来源范围以及原代肝细胞批次间的变异性和功能差异导致药物开发期间预测药物诱导的肝毒性的问题。体外人多能干细胞（hPSC）衍生的肝细胞样细胞有望作为预测药物诱导的肝毒性的工具。一些研究已经报道了将hPSC分化为肝细胞样细胞的有效方法，然而其分化过程耗时、劳动密集、成本密集且不稳定。为了解决这个问题，扩增培养hPSC衍生的肝祖细胞，包括肝干细胞和成肝细胞，其可以自我更新并分化为肝细胞，作为肝细胞的来源应该是有价值的。然而，hPSC衍生的肝祖细胞的扩增机制尚未完全了解。在这项研究中，为了分离hPSC衍生的肝祖细胞，我们试图开发无血清生长因子定义的培养条件，使用定义的组件。我们的培养条件能够分离和生长hPSC衍生的肝祖细胞，其可以通过成肝细胞样细胞分化为肝细胞样细胞。我们已经证实，通过我们的方法制备的肝细胞样细胞能够增加细胞色素P450酶的基因表达后，遇到利福平，苯巴比妥，或奥美拉唑。在确定的培养条件下分离和扩增hPSC衍生的肝祖细胞在检测外源性因素对肝谱系分化的准确影响、理解肝祖细胞自我更新能力的潜在机制以及稳定地供应功能性肝细胞方面具有优势。
Limited growth potential, narrow ranges of sources, and difference in variability and functions from batch to batch of primary hepatocytes cause a problem for predicting drug-induced hepatotoxicity during drug development. Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells in vitro are expected as a tool for predicting drug-induced hepatotoxicity. Several studies have already reported efficient methods for differentiating hPSCs into hepatocyte-like cells, however its differentiation process is time-consuming, labor-intensive, cost-intensive, and unstable. In order to solve this problem, expansion culture for hPSC-derived hepatic progenitor cells, including hepatic stem cells and hepatoblasts which can self-renewal and differentiate into hepatocytes should be valuable as a source of hepatocytes. However, the mechanisms of the expansion of hPSC-derived hepatic progenitor cells are not yet fully understood. In this study, to isolate hPSC-derived hepatic progenitor cells, we tried to develop serum-free growth factor defined culture conditions using defined components. Our culture conditions were able to isolate and grow hPSC-derived hepatic progenitor cells which could differentiate into hepatocyte-like cells through hepatoblast-like cells. We have confirmed that the hepatocyte-like cells prepared by our methods were able to increase gene expression of cytochrome P450 enzymes upon encountering rifampicin, phenobarbital, or omeprazole. The isolation and expansion of hPSC-derived hepatic progenitor cells in defined culture conditions should have advantages in terms of detecting accurate effects of exogenous factors on hepatic lineage differentiation, understanding mechanisms underlying self-renewal ability of hepatic progenitor cells, and stably supplying functional hepatic cells.