诱导大鼠骨髓间充质干细胞分化为肝样细胞的实验研究

目的：探讨肝细胞生长因子（HGF）和粒细胞集落刺激因子（G-CSF）诱导骨髓间充质干细胞（BM-MSCs）分化为肝样细胞的可行性。方法：经骨髓分离BM-MSCs并鉴定,将细胞按以下分组进行成肝诱导：G0组（阴性对照）、G1组（20ng·mL^-1 HGF）、G2组（20ng·mL^-1 HGF＋5ng·mL^-1 G-CSF）、G3组（20ng·mL^-1 HGF＋10ng·mL^-1 G-CSF）、G4组（20ng·mL^-1 HGF＋20ng·mL^-1 G-CSF）,PAS染色检测肝细胞合成糖原功能,利用RT-PCR检测分化肝细胞的甲胎蛋白（AFP）和白蛋白（ALB）表达。结果：G1～G4组BM-MSCs被诱导后呈现肝样细胞形态改变,以G3、G4明显,G0组细胞无类似变化。G1～G4组诱导第7、14天的细胞PAS染色阳性,G0组呈阴性染色;同一时间点,G1与G2、G3与G4比较,差异无统计学意义（P〉0.05）;而G3、G4与G2比较,G3、G4组阳性染色率较高（P〈0.05）。G1～G4组中,诱导AFP于第7天均表达AFP,第14天表达降低,第21天无表达（均P〈0.05）。ALB在第14天出现表达,第21天表达增多,呈上升趋势（P〈0.05）。G2与G1、G4与G3之同比较,AFP、ALB灰度比值差异无统计学意义（P〉0.05）;但G4、G3与G2比较,同一时间点间G4、G3表达水平较高（P〈0.05）。结论：应用HGF和G-CSF能诱导BM-MSCs向肝样细胞分化。但G-CSF需要达到合适浓度才能与HGF发挥协同作用。

Hepatic Differentiation of Bone Marrow Mesenchymal Stem Cells in Vitro

Objective:To explore the feasibility of differentiation of rat BM-MSCs into hepatocyte-like cells induced by hepatocyte growth factor(HGF) and granulocyte colony stimulating factor (G-CSF) in vitro. Methods: Rat BM-MSCs were collected and identified. And then BM-MSCs were divided into the following groups and induced into hepatocyte-like cells. Go served as a negative control, with no HGF or G-CSF. G1 was a positive control, with 20ng/ml HGF. G2 contained 20 ng/ml HGF and 5 ng/ml G-CSF. G3 was 20 ng/ml HGF ...