Extensively expanded murine-induced hepatic stem cells maintain high-efficient hepatic differentiation potential for repopulation of injured livers
Extensively expanded murine-induced hepatic stem cells maintain high-efficient hepatic differentiation potential for repopulation of injured livers
复制标题
广泛扩增的小鼠诱导肝干细胞维持高效肝分化潜力，促进受损肝脏的再生
DOI：
10.1111/liv.14509
复制
发表时间：
2020
影响因子：
6.7
通讯作者：
Hu Yiping
中科院分区：
文献类型：
--
作者：
Yu Bing;Li Hengyu;Chen Jie;He Zhiying;Sun Haixiang;Yang Guangshun;Shang Changzhen;Wang Xin;Li Chuanjiang;Chen Yajin;Hu Yiping
背景与目的供肝细胞的缺乏限制了肝细胞移植的临床应用.诱导的肝干细胞（iHepSC）具有自我更新和双能分化的能力。在这里，我们研究了iHepSC是否可以广泛地扩展，以及它们是否可以分化成足够的功能肝细胞作为供体用于移植therapy.MethodsMurine广泛扩展的iHepSC（50 - 55代）在化学限定的条件下诱导分化成iHepSC-Heps。iHepSC-Hep被证明具有肝细胞形态学特征和肝细胞功能，包括低密度脂蛋白摄取、糖原储存、CLF分泌、ICG摄取和释放、Alb分泌、尿素合成和代谢相关基因表达。接下来，将iHepSC和iHepSC-Heps分别移植到Fah-/-小鼠中。结果小鼠iHepSCs经广泛扩增后仍保持自我更新和双能分化的能力。iHepSCs体外诱导分化为功能性肝细胞的效率达到72.64%。广泛扩增的iHepSC和iHepSC-Heps的移植导致Fah-/-小鼠中的肝脏植入。iHepSC‐Heps组的Fah‐/‐小鼠受体存活率和肝脏再增殖率分别为50%和20.32 ± 4.58%，iHepSC组分别为33%和10.4 ± 4.3%，iHepSC组的Fah‐/‐小鼠受体存活率和肝脏再增殖率分别为50%和20.32 ± 4.58%，iHepSC组分别为33%和10.4 ± 4.3%。在Fah-/-小鼠中，iHepSC-Heps的移植比iHepSC的移植更有效且更高效。我们的结果表明了一种创新的系统，通过谱系重编程产生的iHepSC的肝分化获得足够的肝细胞。
Background & AimShortage of donor hepatocytes limits hepatocyte transplantation for clinical application. Induced hepatic stem cells (iHepSCs) have capacities of self‐renewal and bipotential differentiations. Here, we investigated whether iHepSCs could be extensively expanded, and whether they could differentiate into sufficient functional hepatocytes as donors for transplantation therapy after their extensive expansions.MethodsMurine extensively expanded iHepSCs (50‐55 passages) were induced to differentiate into iHepSC‐Heps under a chemically defined condition. iHepSC‐Heps were proved for carrying morphological hepatocyte characters and hepatocytic functions including low‐density lipoprotein uptake, glycogen storage, CLF secretion, ICG uptake and release, Alb secretion, urea synthesis and metabolism‐relative gene expressions respectively. Next, both iHepSCs and iHepSC‐Heps were transplanted into Fah‐/‐ mice respectively. Both liver repopulation and alleviation of liver function were compared between two transplantation groups.ResultsMurine iHepSCs still maintained the capacities of self‐renewal and bipotential differentiations after extensive expansion. The efficiency for the functional hepatocyte differentiation from extensively expanded iHepSCs reached to 72.64%. Transplantations of both extensively expanded iHepSCs and iHepSC‐Heps resulted in liver engraftment in Fah‐/‐ mice. Survival rate of Fah‐/‐ mice recipients and level of liver repopulation were 50% and 20.32 ± 4.58% respectively in iHepSC‐Heps group, while 33% and 10.4 ± 4.3% in iHepSCs group.ConclusionsExtensively expanded iHepSCs can efficiently differentiate into hepatocytes in chemical defined medium. Transplantation of iHepSC‐Heps was more effective and more efficient than transplantation of iHepSCs in Fah‐/‐ mice. Our results suggested an innovative system to obtain sufficient hepatocytes through hepatic differentiation of iHepSCs generated by lineage reprogramming.