Najimi Mustapha, Khuu Dung Ngoc, Lysy Philippe Antoine, Jazouli Nawal, Abarca Jorge, Sempoux Christine, Sokal Etienne Marc
Université catholique de Louvain, Laboratory of Pediatric Hepatology & Cell Therapy, B-1200 Brussels, Belgium.
Cell Transplant. 2007;16(7):717-28. doi: 10.3727/000000007783465154.
It is currently accepted that adult tissues may develop and maintain their own stem cell pools. Because of their higher safety profile, adult stem cells may represent an ideal candidate cell source to be used for liver cell therapies. We therefore evaluated the differentiation potential of mesenchymal-like cells isolated from adult human livers. Mesenchymal-like cells were isolated from enzymatically digested adult human liver and expanded in vitro. Cell characterization was performed using flow cytometry, RT-PCR, and immunofluorescence, whereas the differentiation potential was evaluated both in vitro after incubation with specific media and in vivo after intrasplenic transplantation of uPA(+/+)-SCID and SCID mice. Adult-derived human liver mesenchymal-like cells expressed both hepatic and mesenchymal markers among which albumin, CYP3A4, vimentin, and alpha-smooth muscle actin. In vitro differentiation studies demonstrated that these mesenchymal-like cells are preferentially determined to differentiate into hepatocyte-like cells. Ten weeks following intrasplenic transplantation into uPA(+/+)-SCID mice, recipient livers showed the presence of human hepatocytic cell nodules positive for human albumin, prealbumin, and alpha-fetoprotein. In SCID transplanted liver mice, human hepatocyte-like cells were mostly found near vascular structures 56 days posttransplantation. In conclusion, the ability of isolated adult-derived liver mesenchymal stem-like cells to proliferate and differentiate into hepatocyte-like cells both in vitro and in vivo leads to propose them as an attractive expandable cell source for stem cell therapy in human liver diseases.
目前已公认成体组织可发育并维持自身的干细胞库。由于成体干细胞具有更高的安全性，它们可能是用于肝细胞治疗的理想候选细胞来源。因此，我们评估了从成人肝脏分离的间充质样细胞的分化潜能。间充质样细胞从经酶消化的成人肝脏中分离出来并在体外扩增。使用流式细胞术、逆转录-聚合酶链反应（RT-PCR）和免疫荧光进行细胞表征，而分化潜能则在与特定培养基孵育后进行体外评估，并在uPA(+/+)-SCID和SCID小鼠脾内移植后进行体内评估。源自成人的人肝脏间充质样细胞表达肝脏和间充质标志物，其中包括白蛋白、CYP3A4、波形蛋白和α-平滑肌肌动蛋白。体外分化研究表明，这些间充质样细胞优先被确定分化为肝细胞样细胞。在脾内移植到uPA(+/+)-SCID小鼠10周后，受体肝脏显示存在对人白蛋白、前白蛋白和甲胎蛋白呈阳性的人肝细胞结节。在移植肝脏的SCID小鼠中，移植后56天大多在血管结构附近发现人肝细胞样细胞。总之，分离的源自成人的肝脏间充质干细胞样细胞在体外和体内增殖并分化为肝细胞样细胞的能力，使得它们成为人类肝脏疾病干细胞治疗中一种有吸引力的可扩增细胞来源。
