两种不同培养基对体外培养脊髓神经元的影响

目的建立体外稳定获取大数量及高纯度脊髓神经元的培养方案，比较当前国内外最常用的无血清培养基（Neurobasal+B27，NCM）与传统含血清培养基（DMEM/F12+10%胎牛血清+5%马血清,SCM）对体外培养脊髓神经元生长状态的影响。方法取E14 15?d Wistar大鼠的胚胎脊髓组织行胰酶消化获取脊髓神经元的单细胞悬液，分别用NCM与SCM进行培养，差速贴壁法及Ara C干预纯化培养的神经元。于接种后第1、2、3、4、5、6、7?d倒置相差显微镜下观察细胞生长状态的变化，并用免疫荧光细胞化学法比较两种培养基培养下获取的神经元纯度。结果采用NCM培养的脊髓神经元生长良好，对环境变化如换液及加入Ara C更为耐受；NCM培养的神经元的纯度为(87.70±8.70)%，SCM培养的神经元纯度为（78.61±7.00）%，前者纯度更高（P=0.019）。结论NCM较之SCM更加适合脊髓神经元的体外培养。

Influence of two media on spinal neurons in vitro

Objective To establish a protocol for harvesting a large number of spinal cord neurons and to compare the effects of two media,NCM(Neurobasal+B27)and SCM (DMEM/F12+10% fetal bovium serum+5% donor equine serum) on spinal neurons cultured in vitro. Methods Embryonic spinal cords of the E Wistar rats on day 14 to 15 were extracted and digested with 0.05% trypsin. Neurons were cultured by the two media for 1 week in vitro. Both physical and pharmacological methods were used to purify the neurons. An inverted ph...