| 摘要: |
| 在无菌条件下抽取成体小鼠骨髓，在含体积分数1% B27,20 ng/mL EGF和10 ng/mL bFGF的F12 DMEM(1∶1) 培养基中，于37 ℃、50 mL/L CO2饱和湿度下常规培养，对培养的细胞进行nestin免疫荧光染色检测，并将pEGFP-N2基因在80 V、80 μs条件下采用电击法转化导入神经干细胞中。结果表明,培养7 d后，有大部分细胞聚集成团，并大量增殖;Nestin免疫荧光染色后呈阳性;导入pEGFP-N2基因的神经干细胞培养后在荧光显微镜下可观察到绿色荧光。表明从成体小鼠获得的骨髓，在该试验培养条件下有神经干细胞生成，并且外源标记基因pEGFP-N2在其细胞中实现了表达。 |
| 关键词: 成体小鼠；骨髓 神经干细胞 分离培养 |
| DOI： |
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| 基金项目:国家“973”重点基础研究项目(G199905400) |
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| Isolation and culture of neural stem cellsfrom bone marrow of adult mouse |
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| Abstract: |
| The bone marrow was obtained from the femur of the adult mouse under the aseptic condition,cultured under 37 ℃ and 50 mL/L CO2 saturated humidity in the F12-DMEM (1∶1) medium,which contains 1% B27,20 ng/mL EGF and 10 ng/ML bFGF.Immunofluorescence stain with Nestin of the separated cells was tested,and pEGFP-N2 was induced in NSCs under condition of 80 V,80 μs by electrotransformation.As a result,the cells aggregate and greatly proliferate in number after one week of culture,immunofluorescence stain with nestin of the isolated cells shows positive result,the NSCs induced with pEGFP-N2 are observed green fluorescence under the fluorescencemicroscope.The result indicates that the bone marrow obtained from the adult mouse in present culture condition contains NSCs,in which exogenetic gene pEGFP-N2 is expressed.This method can be a new way of obtaining NSCs and the foundation of inducing correlated gene to treat neurodegeneration diseases. |
| Key words: adult mouse bone marrow neural stem cells isolation and culture |
