人骨髓间充质干细胞体外分化为雪旺样细胞的方法

目的 :建立人骨髓间充质干细胞体外向雪旺样细胞定向诱导分化的方法。方法 :取健康人骨髓血标本 ,利用percoll(密度为 1.0 73 g·ml-1)分离骨髓单个核细胞 ,在DMEM LG + 10 % (体积分数 )FBS中培养 ,通过流式细胞术对培养细胞进行表型测定 ,对第 2、3、5、8代间充质干细胞进行定向诱导分化 ,用单抗S 10 0、神经胶质纤维酸性蛋白 (glialfibrillargacidicprotein ,GFAP) ,通过免疫组化方法对诱导的细胞进行鉴定。 结果 :经 percoll分离的间充质干细胞可传 16代 ,细胞数增加了大约 6× 10 7倍。流式细胞术结果显示 :percoll分离出来的未经培养的细胞 ,其CD2 9+CD4 4 +CD3 4 -CD4 5 -细胞数为 3 2 .4 7%± 3 .4 9% ,而培养后贴壁细胞中CD2 9+CD4 4 +CD3 4 -CD4 5 -细胞数为 94 .3 8%± 1.5 0 %。诱导后免疫组化染色结果显示 :由 β ME +bFGF预诱导、β ME +bFGF诱导的S 10 0阳性细胞最多 ,可达 90 %± 4 % ,GFAP阳性细胞为 2 1%± 5 %。结论 :人骨髓中间充质干细胞能定向诱导分化成雪旺样细胞。

Method of differentiation of adult humen bone marrow mesenchymal stem cell into Schwann-like cells in vitro

OBJECTIVE: To establish a method of committedly induced adult bone marrow mesenchymal stem cells differentiating into schwann' s-like cells in vitro. METHODS: Adult human bone marrow mononuclear cells were separated by gradient centrifugation on Percoll(density 1.073 g.ml-1). The cells were incubated in low-glucose DMEM with 10% fetal bovine serum. Cell surface antigenic features were analyzed by flow cytometry technique. Passages 2, 3, 5 and 8 of MSCs were committedly induced to differentiate into Schwann-like cells. S-100, GFAP was detected by immunohistochemistry staining. RESULTS: The mesenchymal stem cells separated by percoll were expanded for more than 16 passages, increased about 6 x 10(7) folds. Flow cytometry results showed that: the CD29+ CD44+ CD34- CD45- cells separated from percoll immediately were 32.47% +/- 3.49%; The CD29+ CD44+ CD34- CD45- cells incubated in DMEM-LG with 10% fetal bovine serum were around 94.38% +/- 1.50%. Immunohistochemistry staining results indicated that induced with beta-ME + bFGF S-100 positive cells were 90% +/- 4%, while about 21% +/- 5% cells expressed GFAP. CONCLUSION: MSCs in human adult bone marrow are capable of being committedly induced to differentiate into Schwann-like cells in vitro.