Successful induction of human chemically induced liver progenitors with small molecules from damaged liver
Successful induction of human chemically induced liver progenitors with small molecules from damaged liver
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DOI：
10.1007/s00535-022-01869-5
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发表时间：
2022-03-16
影响因子：
6.3
通讯作者：
Eguchi, Susumu
中科院分区：
文献类型：
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作者：
Miyoshi, Takayuki;Hidaka, Masaaki;Eguchi, Susumu
背景成熟肝细胞产生的小分子诱导的人化学诱导肝祖细胞（human chemical induced liver progenitor，hCLiP）可以潜在地克服肝细胞移植相关的问题，如移植排斥或供体短缺。然而，据我们所知，还没有研究探索从受损肝脏中的成熟肝细胞（MH）诱导hCLiP，指示用于肝移植。方法收集我科86例患者手术切除的肝组织，包括损伤的肝组织，采用胶原酶灌注法分离肝细胞。将从这86名供体中的33名供体中分离的肝细胞在含有Y-27632（ROCK抑制剂）、A-83-01（TGF-β I型受体抑制剂）和CHIR 99021（GSK-3抑制剂）的YAC培养基中培养以诱导hCLiP，并评估其功能。结果无论肝纤维化分类如何，均分离出肝细胞（活力：F0，1：87.2 +/- 13.2%; F2，3：87.8 +/- 13.1%; F4：86.3 +/- 4.2%）。在YAC培养基中培养的大多数肝细胞获得肝祖细胞（LPC）基因。重编程前，hCLiP中MH标志物（ALB、HNF 4 α、G6 PC和CYP 1A 2）的表达低于MH。逆转录-聚合酶链反应显示hCLiP标记物（例如，EpCAM、SOX 9、CK 19和CD 133）在LPC中的表达高于MH。此外，hCLiPs具有分化为肝细胞的能力，并作为成熟肝细胞移植到肝脏表面。结论肝细胞可从损伤肝脏中分离得到。此外，hCLiP可以从分离自受损肝脏的肝细胞获得，并且可以在体外分化成MH。自体hCLiP可以潜在地移植而没有肿瘤发生并重塑受损的肝脏。
Background Human chemically induced liver progenitors (hCLiP) induced by small molecules produced by mature hepatocytes can potentially overcome issues related to hepatocyte transplantation, such as graft rejection or donor shortage. However, to our knowledge, no studies have explored the induction of hCLiP from mature hepatocytes (MHs) in damaged liver, indicated for liver transplantation. Methods Liver tissues were collected from surgically resected livers, including damaged livers, of 86 patients at our department, and hepatocytes were isolated using the collagenase perfusion method. Hepatocytes isolated from 33 of these 86 donors were cultured in YAC medium containing Y-27632 (ROCK inhibitor), A-83-01 (TGF-beta type I receptor inhibitor), and CHIR99021 (GSK-3 inhibitor) to induce hCLiP, and their functions were assessed. Results Hepatocytes were isolated regardless of the liver fibrosis classifications (viability: F0,1: 87.2 +/- 13.2%; F2,3: 87.8 +/- 13.1%; and F4: 86.3 +/- 4.2%). Most hepatocytes cultured in the YAC medium acquired the liver progenitor cell (LPC) gene. The expression of MH markers (ALB, HNF4 alpha, G6PC, and CYP1A2) was lower in hCLiP than in MHs before reprogramming. Reverse transcription-polymerase chain reaction revealed that hCLiP markers (e.g., EpCAM, SOX9, CK19, and CD133) exhibited higher expression in LPCs than in MHs. Furthermore, hCLiPs had the ability to differentiate into hepatocytes, and were engrafted on the liver surface as mature hepatocytes. Conclusion Hepatocytes could be isolated from damaged liver. Furthermore, hCLiP may be obtained from hepatocytes isolated from damaged liver and may differentiate into MHs in vitro. Autologous hCLiP can potentially be transplanted without tumorigenesis and remodel damaged liver.