感觉性和运动性神经膜细胞的培养研究(英文)

A study of sensory and motor Schwann cell culture in vitro

目的对感觉性和运动性神经来源的神经膜细胞进行培养和鉴定,并通过神经生长因子(NGF)的表达间接地研究两种细胞的差别,探讨对神经特异性再生的影响。方法立体显微镜下对SD乳鼠后根神经和股神经运动支进行取材,经2.5g/L胰蛋白酶+0.3g/LIV型胶原酶联合消化,用高糖型DMEM/F12(含100g/LCS)对感觉神经源性和运动神经源性的神经膜细胞进行培养,并经抗S100荧光组织化学染色鉴定。双抗体夹心间接ELISA法测量两种神经膜细胞培养基中NGF的表达。结果培养的两种神经膜细胞经荧光染色证明均为神经膜细胞,光镜观察并手工计数显示两种细胞纯度均超过95%,未见形态学差异,但NGF的表达量和表达模式差异均有显著性意义(F=45.3681,P=0.000)。结论本实验方法可以获得高纯度的感觉性和运动性神经源性神经膜细胞,两者的生物学功能有差异。

Aim To establish the culture method of sensory and motor Schwann cells for the use of nerve tissue engineering and study their biological differences.Methods Sensory and motor Schwann cells were cultured respectively from dorsal root nerves and femoral nerve's motor rami of 5 day old SD rat.The cultured Schwann cells were identified by anti S100 immuno fluorescent immunohistochemical staining,and the NGF expressions of two kinds of cells in the culture media were measured by Sandwich ELISA. Results Both kinds of cells were proved to be Schwann cells.No obvious difference was observed at morphology under light microscope, but their NGF expressions were different in both quantity and pattern (F=45.3681,P=0.000).Conclusion High purity of sensory and motor Schwann cells needed by nerve tissue engineering can be prepared by this method, and there is biological difference between them.