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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。
Objective To culture and identify neuronal cells derived from sensory and motor neurons, and to study the difference between the two kinds of cells through the expression of nerve growth factor (NGF) and to explore the effect on nerve-specific regeneration. METHODS: The dorsal root ganglion and femoral motor branches of SD neonatal rats were harvested under stereomicroscope. The cells were digested with 2.5g / L trypsin + 0.3g / L collagenase and treated with high glucose DMEM / F12 (containing 100g / L) Sensory neurogenic and motor neurogenic neuronal cells were cultured and identified by anti-S100 fluorescent histochemical staining. Double antibody sandwich indirect ELISA was used to measure the expression of NGF in two kinds of neural cell culture medium. Results Both of the cultured neurofilm cells were proved to be mesenchymal cells by fluorescence staining. The purity of both cells was over 95% by light microscopy and manual counting, and no morphological difference was found. However, the expression and expression pattern of NGF were both different There was significant (F = 45.3681, P = 0.000). Conclusion The experimental method can obtain high-purity sensory and motor neurogenic mesenchymal cells, the biological functions of the two are different.