CAJ | 학술논문

目的探索骨髓问充质干细胞(MSCs)体外跨越分化为肝细胞的诱导体系.方法从单核细胞中分离间充质干细胞,用添加20 ng/ml HGF、10 ng/ml FGF-4、10 ng/ml OSM和10%NBS的IMDM培养液诱导间充质干细胞向肝细胞分化.结果经细胞因子诱导的细胞出现肝细胞样变化,随诱导时间延长体积逐渐增大、形态往上皮样转变,胞浆丰富,出现双核或多核细胞.RT-PCR和免疫荧光检测结果显示,AFP在诱导初期表达,在诱导后期表达下降;CKl8、ALB和TAT的表达与诱导时间呈线性关系.第20天达到表达高峰;诱导20 d后免疫荧光检测,AFP、CKl8、ALB和TAT均为阳性;诱导20 d细胞的PAS糖原合成反应呈阳性,即具备糖原合成和储存的肝细胞特有功能.结论 HGF、FGF-4和OSM三种细胞因子组合,可诱导小鼠间充质干细胞向肝细胞分化,该结果为间充质干细胞跨越分化肝细胞的分子机制探讨和临床应用打下了基础.
목적 탐색골수문충질간세포(MSCs)체외과월분화위간세포적유도체계.방법 종단핵세포중분리간충질간세포,용첨가20 ng/ml HGF、10 ng/ml FGF-4、10 ng/ml OSM화10%NBS적IMDM배양액유도간충질간세포향간세포분화.결과 경세포인자유도적세포출현간세포양변화,수유도시간연장체적축점증대、형태왕상피양전변,포장봉부,출현쌍핵혹다핵세포.RT-PCR화면역형광검측결과현시,AFP재유도초기표체,재유도후기표체하강;CKl8、ALB화TAT적표체여유도시간정선성관계.제20천체도표체고봉;유도20 d후면역형광검측,AFP、CKl8、ALB화TAT균위양성;유도20 d세포적PAS당원합성반응정양성,즉구비당원합성화저존적간세포특유공능.결론 HGF、FGF-4화OSM삼충세포인자조합,가유도소서간충질간세포향간세포분화,해결과위간충질간세포과월분화간세포적분자궤제탐토화림상응용타하료기출.
Objective To establish an effective method for inducing mouse bone marrow mesen-ehymal stem cells (MSCs) into hepatocytes. Methods Isolated MSCs were selected by plastic adher-ence, differentiated in Iscove's Modified Dulbecco's medium supplemented with 10% new bovine ser-um (NBS), 20 ng/mL hepatocyte growth factor (HGF), 10 ng/mL fibroblast growth factor-4 (FGF-4) and 10 ng/ml oncostatin m (OSM) for 20 days'induction. The medium was changed every 4 days.Results When MSCs were cultured with HGF, FGF-4 and OSM, cuboidal morphology, which was a characteristic of hepatocytes, was observed, and the differentiated ceils also expressed marker genes specific to liver cells in a time-dependent manner, a-fetoprotein (AFP) was expressed on day 10, and CK18, ALB and TAT were detected on day 20, which was in consistent with the immunofluoresence results. Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes of storing glycogen. Conclusion Mouse MSCs can differentiate into hepatocytes when in-duced by HGF, FGF-4 and OSM.