摘要
目的建立体外视网膜神经节细胞（ｒｅｔｉｎａｌｇａｎｇｌｉｏｎｃｅｌｓ，ＲＧＣｓ）培养和纯化模型。方法出生后１～３天ＳｐｒａｇｕｅＤａｗｌｅｙ（ＳＤ）乳鼠，ＲＧＣｓ在ｂａｓａｌｍｅｄｉｕｍｅａｇｌｅ（ＢＭＥ）培养基中培养，相差显微镜下观察细胞生长规律。羊抗鼠单克隆抗体ＦＩＴＣＴｈｙ１．１鉴定ＲＧＣｓ，并通过Ｔｈｙ１．１单抗进行ＲＧＣｓ的分离纯化。四唑盐（ｍｏｎｏｔｅｔｒａｚｏｌｉｕｍ，ＭＴＴ）微量比色法检测细胞活性。结果较高的细胞接种密度或加入顶盖提取液（ｔｅｃｔａｌｅｘｔｒａｃｔ，Ｔｅ）均有利于混合培养的ＲＧＣｓ生长存活。ＭＴＴ检测法显示加Ｔｅ培养的实验组其吸光度（Ａ）值较对照组增加了３倍；在ＲＧＣｓ培养２天时，按细胞接种密度为１×１０３个／ｍｍ２的培养组较５００个／ｍｍ２的培养组，有突起的细胞数多３倍以上。ＲＧＣｓ经Ｔｈｙ１．１单抗纯化，其纯化率约达９５％。在无神经营养因子的条件下，纯化培养的ＲＧＣｓ于４８ｈ后很少存活。而混合培养的ＲＧＣｓ存活时间可持续２或３周。结论在一定条件下的ＲＧＣｓ能够于体外培养存活，其生长存活能力依赖于靶组织及其它视网膜细胞的营养作用。采用Ｔｈｙ１．１单克隆抗体纯化的方法可?
Objective To establish a cell
line and purification model of retinal ganglion cells (RGCs) in vitro. Method RGCs from Sprague
Dawley neonatal rats (postnatal 13 days) were cultured in basal medium eagle (BME) basal
medium. The growth regularity of RGCs in vitro was observed under phasecontrast
microscope. RGCs were purified by Thy 1.1 with FITC antibody and detected under fluorescent
microscope and phasecontrast microscope. Results Higher density of retinal cells and tectal
extract facilitate cultured RGCs to survive. The purification rate of retinal ganglion cells in the
experiment arrived at 95 percent. Conclusion Cytokine and trophic factors from other cells in
the retina and tectal extract can promote RGCs to survive, and they can be purified by Thy 1.1
antibody.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
1999年第3期190-193,10,共5页
Chinese Journal of Ophthalmology
基金
国家自然科学基金
广东省科委重点攻关项目基金
广东省中医药管理局基金
