视网膜神经节细胞的培养

Retinal Ganglion Cells in Culture

视网膜神经节细胞（ｒｅｔｉｎａｌｇａｎｇｌｉｏｎｃｅｌｓ，ＲＧＣｓ）是青光眼性视神经病变的主要损害细胞。近年来发展起来的ＲＧＣ培养技术为揭示其损害机制提供了１条有效途径。因此，我们需了解获得ＲＧＣｓ悬液最普遍的视网膜完全消化法及部分消化剥脱法，以及如何对ＲＧＣｓ进行分离、培养及鉴定技术。在作ＲＧＣｓ培养前需对培养皿、培养瓶等进行处理，常覆以聚－Ｌ－赖酸或预先培养的单层胶质细胞。在细胞鉴定方面，采用荧光逆行标记和Ｔｈｙ－１单克隆抗体免疫组化及细胞内电记录等方法。Ｔｈｙ－１单克隆抗体、板层素（ｌａｍｉｎｉｎ）、神经生长因子（ｎｅｕｒｏｇｒｏｗｔｈｆａｃｔｏｒｓ，ＮＧＦ）、ａ－纤维母细胞生长因子（ａ－ｆｉｂｒｏｂｌａｓｔｓｇｒｏｗｔｈｆａｃｔｏｒｓ，ａＦＧＦ）等，对培养ＲＧＣｓ有一定影响。

Retinal ganglion cells(RGCs) are the mainly pathological cells in glaucoma.The RGCs culture technique,which developed in the past few years,provides an effective mode for illustrating its pathological mechanism.The article demonstrated the dissociation,culture and identification technique of RGCs.Recently,the most popular methods for making RGCs suspension solution are the entire digestion method and partial digestion peeling method.It is necessary for RGCs culture to cover a substratum on the culture dish,which usually is covered a layer of polyLlysine or preceeded to culture a glial cells layer.The cultural RGCs are identified by fluorescein retrograde mark,Thy1 monoclonal antibody immunocytochemistry method or intracellular electrophysiology method,etc.The cultured RGCs are affected by Thy1 monoclonal antibody,laminin,neuro growth factors(NGF),fibroblast growth factors(FGF),etc.