Neural cell differentiation in vitro from adult human bone marrow mesenchymal stem cells
Neural cell differentiation in vitro from adult human bone marrow mesenchymal stem cells
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DOI：
10.1089/scd.2005.14.65
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发表时间：
2005-02-01
影响因子：
4
通讯作者：
Kletzel, M
中科院分区：
文献类型：
--
作者：
Long, XX;Olszewski, M;Kletzel, M
骨髓间充质干细胞是一种能够在体外扩增并分化为非造血细胞的细胞。神经营养细胞因子，如人表皮生长因子（hEGF）和牛成纤维细胞生长因子（bFGF）可以诱导分化为神经细胞（NC）。当骨髓间充质干细胞与hEGF和bFGF共同培养时，观察到神经元特异性标志物Nestin、MAP-2和酪氨酸羟化酶（TH）的RNA表达。我们测试了新的细胞因子组合以产生成熟的NC。收集塑料贴壁细胞，然后当它们与富集的单核细胞层90%汇合时分离。在第3代，MSC在神经分化培养基（在NEUROBASAL培养基中的dbcAMP、IBMX、FGF-8、BDNF、hEGF和bFGF加上B27）中培养。在第6天计数细胞。免疫荧光染色和逆转录酶（RT）-PCR检测神经标志物的表达。在第6天，66%的细胞发育成树突，并呈现典型的神经细胞形态。一些细胞的早期神经标志物巢蛋白和β-微管蛋白III呈阳性。还观察到表达成熟神经元标记物（NF、NeuN、Tau、Nurr 1、GABA、寡树突GalC和胶质GFAP）的细胞。通过将hEGF、bFGF、dbcAMP、IBMX、BDNF和bFGF-8加入到含有B27的神经基础培养基中，将BM MSC定向为早期和成熟的NCs。
Bone marrow ( BM) mesenchymal stem cells (MSCs) are cells capable of expanding and differentiating in vitro into nonhematopoietic cells. Neurotrophic cytokines, such as human epidermal growth factor ( hEGF) and bovine fibroblast growth factor ( bFGF) can induce differentiation into neural cells (NCs). When BM MSCs were cultured with hEGF and bFGF, RNA expression of neuronal specific markers Nestin, MAP-2, and tyrosine hydroxylase (TH) were observed. We tested a new cytokine combination to generate mature NCs. The plastic-adherent cells were collected and then split when they were 90% confluent from an enriched mononuclear cell layer. At passage 3, MSCs were cultured in neural differentiation media ( dbcAMP, IBMX, FGF-8, BDNF, hEGF, and bFGF in NEUROBASAL media plus B27). Cells were counted on day 6. Immunofluorescent staining and reverse transcriptase (RT)-PCR were performed to evaluate the expression of neural markers. On day 6, 66% of cells developed dendrites and presented typical neural cell morphology. Some cells were positive for early neural markers Nestin and beta-tubulin III. Cells expressing mature neuronal markers (NF, NeuN, Tau, Nurr1, GABA, oligodendryte GalC, and glial GFAP) were also seen. By adding hEGF, bFGF, dbcAMP, IBMX, BDNF, and bFGF-8 into NEUROBASAL media plus B27, BM MSCs were directed toward becoming early and mature NCs.