中文摘要：

目的建立以30 ng/ml FGF-4和30 ng/ml OSM为主的诱导培养体系,研究体外骨髓干细胞向肝细胞的转化过程中细胞形态、结构、功能的变化.方法建立以30ng/ml FGF,30ng/mlOSM为主的小鼠骨髓干细胞诱导培养体系.观察细胞形态和数量的变化;RT-PCR、Western blot、免疫荧光染色检测肝细胞特异性标记物的表达;通过PAS糖原染色、白蛋白ELISA、尿素合成试验,检测细胞的合成和代谢功能.结果诱导12 d,观察到多极性的肝细胞样细胞,且逐渐增多、集落不断增大.在诱导过程中,不同的时间点可以分别检测到HNF-3βmRNA、ALBmRNA、CK18mRNA、TTRmRNA、G6-PasemRNA和TATmRNA的表达.诱导21 d,细胞表达HNF-3β、ALB、CK18蛋白;免疫荧光定位示ALB表达于胞浆和胞膜,而CK18表达于胞浆.诱导21 d,细胞胞浆内可见红染的糖原颗粒;在诱导过程中,不同的时间点检测到细胞分泌ALB、合成尿素的量的变化.结论我们建立的以30ng/ml FGF、30 ng/ml OSM为主的诱导培养体系,可以有效地促进骨髓干细胞体外定向转化为肝细胞.

英文摘要：

Objective To establish the directed differentiation media including 30 ng/ml FGF-4 and 30ng/ml OSM and reveal the differentiation of bone marrow cells （BMSCs） into hepatocytes by morphological examination and determination of rnRNA expression, protein expression and cell function. Methods The mouse BMSCs were cultured in the directed differentiation media including FGF-4 and OSM. Morphology of the hepatocytes form directed differentiation of BMSCs were identified. RNA expression of the hepatocytes was tested by RT-PCR and the protein expression by Western blotting and immunofluorescent assay. Synthetic and metabolic functions of the hepatocytes were investigated by albumin ELISA, periodic acid Shill staining and urea assay. Results Some epitheliumlike cells or polygonal cells appeared and increased in the course of the cell directed differentiation. HNF-313, ALB, CK18, TTR, G-6-Pase and TAT mRNA were expressed in the course of the directed differentiation. The directed differentiated cells expressed HNF-313, ALB and CK18 on day 21. ALB was found in the cytoplasm and cell membrane while CK18 scattered in the cytoplasm. The differentiated cells produced albumin and synthesized urea in a time-dependent manner. They could also synthesize glycogen. Conclusions The differentiation media including FGF-4 and OSM is effective to differentiate mouse BMSCs into hepatoeytes.