Paracrine signals from mesenchymal cell populations govern the expansion and differentiation of human hepatic stem cells to adult liver fates.
Paracrine signals from mesenchymal cell populations govern the expansion and differentiation of human hepatic stem cells to adult liver fates.
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DOI：
10.1002/hep.23829
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发表时间：
2010-10
期刊：
影响因子：
13.5
通讯作者：
Reid, Lola M.
中科院分区：
文献类型：
--
作者：
Wang, Yunfang;Yao, Hsin-Lei;Cui, Cal-Bin;Wauthier, Eliane;Barbier, Claire;Costello, Martin J.;Moss, Nicholas;Yamauchi, Mitsuo;Sricholpech, Marnisa;Gerber, David;Loboa, Elizabeth G.;Reid, Lola M.
在已知条件下，胚胎或定向干细胞群向成体肝脏细胞分化会产生具有部分而非全部成体特异性基因的细胞，一个或多个基因的调控异常，且实验结果存在差异。我们验证了这样一个假设：由新鲜分离的、谱系依赖的间充质细胞群产生的一系列信号在驱动人类肝干细胞（hHpSCs）向成体肝脏细胞分化时会产生更高的效率和可重复性。通过免疫选择技术纯化的肝脏来源间充质细胞亚群包括：1）成血管细胞；2）成熟内皮细胞；3）肝星状细胞前体；4）成熟星状细胞（周细胞）和5）肌成纤维细胞。在我们为短期培养所开发的完全确定（无血清）的条件下，将这些亚群中每种细胞的新鲜免疫选择细胞建立在原代培养中，并将它们用作hHpSCs的饲养层细胞。成血管细胞饲养层导致自我复制；星状细胞前体使细胞谱系限制为肝母细胞；成熟内皮细胞促使分化为肝细胞；成熟星状细胞和/或肌成纤维细胞导致分化为胆管细胞。通过生化、免疫组织化学和定量逆转录聚合酶链反应（qRT - PCR）分析确定了不同饲养层细胞产生的旁分泌信号，然后在单层和三维培养中使用这些信号替代饲养层细胞，以从hHpSCs中引发所需的生物学反应。确定的旁分泌信号被证明能够使hHpSCs产生可重复的反应，并使其分化为完全成熟且具有功能的实质细胞。
来自特定间充质细胞群的旁分泌信号对于将干细胞群调控为特定的成体命运非常重要，这一发现对于基础和临床研究以及工业研究都具有重要意义。
Differentiation of embryonic or determined stem cell populations to adult liver fates under known conditions yields cells with some but not other adult-specific genes, aberrant regulation of one or more genes, and variation in the results from experiment to experiment. We tested the hypothesis that sets of signals produced by freshly isolated, lineage-dependent mesenchymal cell populations would yield greater efficiency and reproducibility in driving differentiation of human hepatic stem cells (hHpSCs) to adult liver fates. Subpopulations of liver-derived mesenchymal cells, purified by immunoselection technologies, included 1) angioblasts; 2) mature endothelia; 3) hepatic stellate cell precursors; 4) mature stellate cells (pericytes) and 5) myofibroblasts. Freshly immunoselected cells of each of these subpopulations were established in primary cultures under wholly defined (serum-free) conditions that we developed for short-term cultures and used them as feeders with hHpSCs. Feeders of angioblasts yielded self-replication; stellate cell precursors caused lineage restriction to hepatoblasts; mature endothelia produced differentiation to hepatocytes; and mature stellate cells and/or myofibroblasts resulted in differentiation to cholangiocytes. Paracrine signals, produced by the different feeders, were identified by biochemical, immunohistochemical, and qRT-PCR analyses and then those signals were used to replace the feeders in monolayer and 3-D cultures to elicit the desired biological responses from the hHpSCs. The defined paracrine signals proved able to yield reproducible responses from the hHpSCs and to permit differentiation to fully mature and functional parenchymal cells. paracrine signals from defined mesenchymal cell populations are important for regulation of stem cell populations to specific adult fates, findings of importance for basic and clinical research as well as industrial investigations.
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