Hepatic stem cells with self-renewal and liver repopulation potential are harbored in CDCP1-positive subpopulations of human fetal liver cells.
Hepatic stem cells with self-renewal and liver repopulation potential are harbored in CDCP1-positive subpopulations of human fetal liver cells.
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人胎儿肝细胞CDCP1阳性亚群中蕴藏着具有自我更新和肝脏再生潜力的肝干细胞
DOI：
10.1186/s13287-017-0747-3
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发表时间：
2018-02-05
影响因子：
7.5
通讯作者：
Taniguchi H
中科院分区：
文献类型：
--
作者：
Zhang RR;Zheng YW;Li B;Nie YZ;Ueno Y;Tsuchida T;Taniguchi H
背景成熟的人肝细胞在肝病的临床前研究和治疗中至关重要，但在体外难以操作和扩增。肝干细胞（HpSC）可能是用于细胞治疗和疾病建模的功能性肝细胞的替代来源。由于这些细胞在再生医学中发挥着重要作用，因此确定用于分离这些细胞的特异性标记物以及它们是否有助于肝再生的精确表征仍有待于显示。MethodIn这项研究中，人HpSCs通过流式细胞术使用CDCP 1，CD 90和CD 66抗体从人原代胎肝细胞（FLC）中分离出来。将分离的CDCP 1 + CD 90 + CD 66-HpSC在用IV型胶原包被的培养皿上在补充有FBS、人γ-胰岛素、烟酰胺、地塞米松和l-谷氨酰胺的DMEM营养混合物F-12 Ham中培养至少2周，并通过转录组学谱分析、定量实时PCR、免疫细胞化学、结果纯化的CDCP 1 + CD 90 + CD 66-亚群具有克隆扩增和自我更新能力，在含有不同肝细胞和胆管细胞的单细胞来源的集落中进一步鉴定出双能能力。此外，体内肝脏再增殖实验表明，人CDCP 1 + CD 90 + CD 66-HpSCs再增殖超过90%的小鼠肝脏，并分化为具有药物代谢活性的功能性肝细胞。表明CDCP 1标记物可用于鉴定和分离HpSC，用于肝脏疾病的进一步细胞治疗。
BackgroundMature human hepatocytes are critical in preclinical research and therapy for liver disease, but are difficult to manipulate and expand in vitro. Hepatic stem cells (HpSCs) may be an alternative source of functional hepatocytes for cell therapy and disease modeling. Since these cells play an import role in regenerative medicine, the precise characterization that determines specific markers used to isolate these cells as well as whether they contribute to liver regeneration still remain to be shown.MethodIn this study, human HpSCs were isolated from human primary fetal liver cells (FLCs) by flow cytometry using CDCP1, CD90, and CD66 antibodies. The isolated CDCP1+CD90+CD66–HpSCs were cultured on dishes coated with type IV collagen in DMEM nutrient mixture F-12 Ham supplemented with FBS, human γ-insulin, nicotinamide, dexamethasone, andl-glutamine for at least 2 weeks, and were characterized by transcriptomic profiling, quantitative real-time PCR, immunocytochemistry, and in-vivo transplantation.ResultsThe purified CDCP1+CD90+CD66–subpopulation exhibited clonal expansion and self-renewal capability, and bipotential capacity was further identified in single cell-derived colonies containing distinct hepatocytes and cholangiocytes. Moreover, in-vivo liver repopulation assays demonstrated that human CDCP1+CD90+CD66–HpSCs repopulated over 90% of the mouse liver and differentiated into functional hepatocytes with drug metabolism activity.ConclusionsWe identified a human hepatic stem/progenitor population in the CDCP1+CD90+CD66–subpopulation in human FLCs, indicating CDCP1 marker could potentially be utilized to identify and isolate HpSCs for further cytotherapy of liver disease.
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影响因子：
3.6
作者：
Li J;Xin J;Zhang L;Wu J;Jiang L;Zhou Q;Li J;Guo J;Cao H;Li L
通讯作者：
Li L
影响因子：
46.9
作者：
Azuma, Hisaya;Paulk, Nicole;Grompe, Markus
通讯作者：
Grompe, Markus
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
--
作者：
Jones, EA;Kinsey, SE;McGonagle, D
通讯作者：
McGonagle, D
影响因子：
4.2
作者：
Grompe, Markus
通讯作者：
Grompe, Markus