【摘要】 目的:观察不同培养环境对小鼠视网膜神经细胞的影响。方法： 分别培养新生小鼠的视网膜神经细胞和色素上皮细胞，并纯化培养视网膜神经节细胞，将纯化和未纯化的视网膜神经细胞分别与视网膜色素上皮细胞接触和分层共培养，相差显微镜观察视网膜神经细胞在不同培养模式中的表现。结果：混合培养的视网膜神经细胞较纯化培养的神经节细胞生存时间长，突触联系多。与视网膜色素上皮细胞共培养时，神经细胞的存活时间进一步延长，突触联系增多，但是神经细胞在与视网膜色素上皮细胞分层和接触共培养的两种模式中，显示出了不同的细胞形态。结论：视网膜神经胶质细胞和视网膜色素上皮细胞不但对神经节细胞具有滋养作用，细胞间的立体结构关系可能还影响着细胞的成熟分化。
【关键词】 视网膜神经节细胞;视网膜色素上皮细胞;共培养
Appearance of murine retinal neural cell in different culture systems
YuPing Zheng, ZhaoHui Feng, LuYan Zhang, HuaNing She, XiaoHua Wang, QuanChen Xiong
Foundation item: National Natural Science Foundation of China (No.30772598)
Department of Ophthalmology, the Second Hospital of Medical College of Xian Jiaotong University, Xian 710004, Shaanxi Province, China
AbstractAIM: To explore the effect of different culture environments on murine retinal neural cells.
METHODS: Neonatal murine retinal neural cells, purified retinal ganglion cells and pigment epithelial cells were cultured. Purified and unpurified retinal ganglion cells were cultured with RPE in contacting or uncontacting modes. Appearance of neural cells was observed under phasecontrast microscope.
RESULTS: Compared with purified ganglion cells, mixed retinal neural cells survived longer and had more synapse. Being cultured with RPE further, both mixed retinal neural cells and purified ganglion cells had longer survive time and more synapse. However, neural cells in contacting mode showed different appearance.
CONCLUSION: In addition to providing nourishment to retinal ganglion cells, retinal glial cells and pigment epithelial cells also affect the differentiation and maturation of retinal neural cells by their stereostructures.
KEYWORDS: retinal ganglion cell; retinal pigment epithelium; coculture
Zheng YP, Feng ZH, Zhang LY, et al. Appearance of murine retinal neural cell in different culture systems. Int J Ophthalmol(Guoji
Yanke Zazhi)2008;8(11):
0引言
视网膜神经细胞，尤其是视网膜神经节细胞( retinal ganglion cells, RGCs)的损伤和死亡，是多种眼底疾病导致失明的共同病变[1]。在本研究中，我们对新生小鼠的视网膜神经细胞在不同的培养模式中进行培养，希望能找到一种与体内环境非常类似的视网膜神经细胞培养体系，从而了解细胞在正常和病理状态下的生物学表现，为遏制视网膜神经病变提供基础。
1材料和方法
1.1材料 C578L/6N小鼠,生后1d和出生后2wk的乳鼠,由本校实验动物中心提供。DMEM/F12培养液（Gibco公司），胎牛血清（Hyclone 公司）。大鼠抗小鼠Thy1 单克隆抗体（Santa Cruz 公司），小鼠抗小鼠 cytokeratin多克隆抗体（博士德公司），羊抗鼠IgGTR（Santa Cruz 公司）。
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