小鼠骨髓间充质干细胞与脑胶质细胞及基底膜细胞共培养后向毛细胞样细胞分化

Differentiation of bone marrow mesenchymal stem cells into hair like cells by co-culture with astrocytes and basement membrane cells of mice

目的探讨体外建立诱导小鼠骨髓间充质干细胞(MSCs)向毛细胞分化的方法。方法分离培养小鼠骨髓间充质干细胞、脑胶质细胞(ASTs)与基底膜细胞(BAMCs)至第3代。取MSCs与ASTs共培养于DMEM/F12/RA+10%FBS培养基中。7 d后免疫荧光法检测Nestin的表达。所得细胞再与BAMCs共培养于DMEM/F12+10%FBS中。14 d后免疫荧光法检测诱导后细胞中毛细胞的标志物(Math1与MyosinVIIa)的表达。RT-PCR检测毛细胞的标志物(Math1与MyosinVIIa)mRNA的表达。结果经ASTs共培养后的MSCs表达Nestin。Nestin阳性细胞与BAMCs共培养后部分细胞表达Math1和MyosinVIIa的蛋白与mRNA。结论 MSCs经与ASTs及BAMCs共培养后,部分细胞被诱导分化为毛细胞样细胞。

Objective To establish a method of efficient production of hair like cells from bone marrow mesenchymal stem cells（MSCs）in vitro.Methods MSCs、astrocytes（ASTs） and basement membrane cells（BAMCs） were isolated from mice,passed and depurated into third generation.We co-cultured MSCs with astrocytes（ASTs） in DMEM/F12/RA＋10% FBS for 7 days,then checked the surface molecule of Nestin by immunocytochemical analyses.Afterward,These cells were co-cultured with BAMCs for 14 days in DMEM/F12＋10% FBS.Then checked the expression of hair cell markers（MyosinVIIa and Math1）by immunocytochemical analyses.We checked the mRNA expression of hair cell markers（MyosinVIIa and Math1）by RT-PCR.Results MSCs which were co-cultured with ASTs for 7 days expressed Nestin.Then Nestin-positive cells co-cultured with BAMCs for 14 days could express mRNA and protein of hair cell markers（Math1 and MyosinVIIa）.Conclusions MSCs co-cultured with ASTs and BAMCs could be induced to differentiate into hair like cells.