摘要
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.
出处
《基础医学与临床》
CSCD
北大核心
2012年第3期283-290,共8页
Basic and Clinical Medicine
基金
福建省科技厅
科技合作重点项目(2008I 0017)