Hepatocyte growth factor in combination with interleukin-6 to induce the in vitro differentiation of mesenchymal stem cells isolated from human umbilical cord blood into hepatocyte-like cells
- VernacularTitle:白细胞介素6和肝细胞生长因子体外诱导人脐血间充质干细胞向肝样细胞分化
- Author:
Hong YUAN
;
Yuzhuo WU
;
Hong CHEN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: Mesenchymal stem cells (MSCs) isolated from human umbilical cord blood can differentiate into hepatocyte-like cells under a suitable induction condition. The optimal condition is still unclear. This study investigated the feasibility of differentiation of MSCs isolated from human umbilical cord blood into hepatocyte-like cells with hepatocyte growth factor (HGF) and interleukin-6 (IL-6) in vitro to find a new cell source for live tissue engineering. METHODS: The experiment was performed from March to September 2007 at the Central Laboratory of First Hospital of Lanzhou University. ①Three cases of umbilical cord blood were collected from full-term pregnant women. Pregnant women had signed an informed consent. The experiment was approved by Hospital’s Ethical Committee. ②MSCs from umbilical cord blood were separated by density gradient centrifugation and adherent method. Cell surface molecule was measured by flow cytometry. Third passage of cells were divided into four groups and cultured in DMEM medium with HGF (HGF group), IL-6 (IL-6 group), HGF+IL-6 (HGF+IL-6 group) or no growth factor (control group). ③The characteristics of proliferation and growth of MSCs were studied by microscopy and methyl thiazolyl tetrazolium (MTT). The phenotypes of MSCs were identified by flow cytometry and immunohistochemical method. Albumin levels in culture supernatants were determined with enzyme-labeled immunosorbent assay (ELISA). RESULTS: ①Growth and division of adherent cells obtained from human umbilical cord blood were good. The shape of MSCs changed into triangle, polygonal or round on days 21-28 in induction groups. ②Positive staining reaction for alpha fetoprotein (AFP) on day 7, for CK18 on days 21, 28 after induction. Albumin production by induced MSCs increased in a time-dependent manner. The positive percents of every hepatocytic marker were higher in the HGF+IL-6 group than in the IL-6 group and HGF group (P
