脂肪源性干细胞促进大鼠神经施万细胞增殖和分泌功能

Adipose-derived Stem Cells Promote Rat Schwann Cell Proliferation and Secretion of Nerve Regeneration Factors

目的检测脂肪源性干细胞(ADSCs)对大鼠神经施万细胞增殖及分泌功能的影响。方法从SD大鼠腹股沟区脂肪中提取ADSCs,通过流式细胞仪检测表面抗体标记物。获取ADSCs条件培养基,通过ELISA判断ADSCs条件培养基中神经再生相关因子[神经生长因子(NGF)、脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)和血管内皮生长因子(VEGF)]的含量。分别将大鼠神经施万细胞与ADSCs条件培养基及ADSCs共培养,通过MTT观察ADSCs条件培养基及ADSCs对大鼠神经施万细胞增殖的影响,进一步通过Western blotting和实时PCR分别检测ADSCs条件培养基及ADSCs作用于大鼠神经施万细胞后神经再生相关因子蛋白和m RNA水平的改变。结果细胞表面抗体标记物检测发现CD44和CD90阳性,CD31和CD45阴性,证明ADSCs提取成功。ELISA检测发现ADSCs条件培养基中含有大量神经再生相关因子。MTT检测发现ADSCs条件培养基及ADSCs均可以促进大鼠神经施万细胞的增殖。Western blotting和实时PCR结果证明ADSCs条件培养基及ADSCs可以促进大鼠神经施万细胞神经再生相关因子的表达。结论 ADSCs可以促进大鼠神经施万细胞增殖及神经再生相关因子分泌。

Objective To detect the effects of adipose-derived stem cells（ADSCs） on proliferation of rat Schwann cells and secretion of nerve regeneration-related factors from Schwann cells. Methods ADSCs were extracted from Sprague Dawley rats,and surface antibody markers were identified by flow cytometry. ADSC-conditioned medium was obtained and the contents of nerve regeneration-related factors（NGF,BDNF,GDNF,and VEGF） in the medium were determined by enzyme-linked immunosorbent assay. Schwann cells were incubated either with the ADSC-conditioned medium or with ADSCs. The effect of the medium and ADSCs on the proliferation of Schwann cells was observed by MTT. The levels of nerve regeneration-related factors（NGF,BDNF,GDNF,and VEGF） in Schwann cells were detected by Western blotting and real-time polymerase chain reaction. Results The evaluation of cell surface antibody markers revealed CD44 and CD90 positivity and CD31 and CD45 negativity,indicating that the extraction of ADSCs was successful. Enzyme-linked immunosorbent assay revealed that the ADSC-conditioned medium contained an elevated amount of nerve regeneration-related factors. MTT assay showed that the ADSC-conditioned medium and ADSCs can promote the proliferation of Schwann cells. Western blotting and real-time polymerase chain reaction showed that ADSC-conditioned medium and ADSCs could promote the expression of nerve regeneration-related factors in Schwann cells. Conclusion ADSCs can promote Schwann cell proliferation and secretion of nerve regeneration factors.