Wang Jing, Liu Kaiyan
Institute of Hematology, Beijing University People's Hospital, 100044, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2003 Apr 18;35(2):202-6.
To establish a method of committedly induced adult bone marrow mesenchymal stem cells differentiating into schwann' s-like cells in vitro.
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.
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.
MSCs in human adult bone marrow are capable of being committedly induced to differentiate into Schwann-like cells in vitro.
建立一种在体外定向诱导成人骨髓间充质干细胞分化为雪旺氏样细胞的方法。
采用密度为1.073 g/ml的Percoll梯度离心法分离成人骨髓单个核细胞。将细胞接种于含10%胎牛血清的低糖DMEM培养基中培养。采用流式细胞术分析细胞表面抗原特性。取第2、3、5和8代骨髓间充质干细胞定向诱导分化为雪旺氏样细胞，采用免疫组织化学染色检测S-100、胶质纤维酸性蛋白（GFAP）。
经Percoll分离的间充质干细胞可传代培养16代以上，扩增约6×10⁷倍。流式细胞术结果显示：Percoll分离后即刻的CD29⁺CD44⁺CD34⁻CD45⁻细胞占32.47%±3.49%；接种于含10%胎牛血清的低糖DMEM培养基中培养后的CD29⁺CD44⁺CD34⁻CD45⁻细胞约占94.38%±1.50%。免疫组织化学染色结果表明，经β-巯基乙醇（β-ME）+碱性成纤维细胞生长因子（bFGF）诱导后，S-100阳性细胞占90%±4%，而GFAP阳性细胞约占21%±5%。
成人骨髓间充质干细胞在体外能够定向诱导分化为雪旺氏样细胞。
