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The study on the transfection of pEGFP-C2 gene into the neural stem cells derived from the bone marrow stromal cells of rhesus monkey
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROMEDICINE
- Issue:
- 2
- DOI:
- 10.3760/cma.j.issn.1671-8925.2004.02.002
- Key Word:
- 骨髓基质细胞;神经干细胞;EGFP;基因转染;恒河猴
Abstract： 目的探讨恒河猴骨髓基质源神经干细胞进行基因修饰的可行性.方法分离恒河猴骨髓基质细胞(BMSCs),体外培养,bFGF诱导增殖,细胞生长至神经干细胞期时以NucleofectorTM核转染仪行pEGFP-C2转染,倒置荧光显微镜观察基因表达情况,并检测细胞的活力.另外,对同期培养的未转染细胞进行神经干细胞特异性抗原-nestm和CD133抗原的免疫细胞化学检测.结果分离得到的BMSCs能在体外培养液中进行增殖和分化,而在bFGF诱导的情况下,细胞的增殖更为明显;转染pEGFP-C2的细胞于转染后24 h后即表达强绿色荧光蛋白,转染率达30%以上,且细胞的活力基本不受影响:免疫细胞化学检测可见有nestin和CD133抗原在同期培养的未转染细胞内表达.结论灵长类骨髓基质细胞具有向神经干细胞分化、增殖的能力,bFGF能促进细胞的增殖,在神经干细胞培养条件下,可发育分化成神经干细胞;在神经干细胞阶段,应用电转染技术进行神经干细胞基因修饰是可行的,可应用于骨髓基质源神经干细胞的基因治疗领域.
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