兔角膜缘上皮细胞在体外壳聚糖共混膜上的培养及生物学鉴定  

Primary culture and differentiation of rabbit limbal epithelial cells cultured on Chitosan-Collagen/Chondroitin sulfate membrane in vitro

作　　者：丁勇[1] 徐锦堂[1] 陈建苏[1] 陈剑[1] 吴春云[2] 张志雄[3] 周长忍[3] 

机构地区：[1]暨南大学附属第一医院眼科,广东广州510632 [2]暨南大学医学院病原学教研室,广东广州510632 [3]暨南大学生命科学院,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2005年第2期205-209,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省自然科学基金 (0 4 0 10 4 32 ) ;广东省医学科研基金 (A2 0 0 4 338) ;广东省重大科技计划项目 (2 0 0 3A2 0 30 10 4 ) ;国家自然科学基金 (30 3715 19)资助项目

摘　　要：目的:探讨壳聚糖-胶原-硫酸软骨素共混膜作为兔角膜缘上皮细胞体外培养载体的可行性。方法:采用消化培养法,用特殊培养液在壳聚糖-胶原-硫酸软骨素共混膜上培养兔角膜缘上皮细胞,第7d应用免疫组化方法对培养细胞作生物学鉴别并在扫描电子显微镜下观察细胞形态。结果:上皮细胞AE1/AE5单克隆抗体免疫组化阳性,培养第7d的上皮细胞有多种细胞形态,细胞胞体饱满,相互通过突起连接,表面可见微绒毛。结论:兔角膜缘上皮细胞可在壳聚糖-胶原-硫酸软骨素共混膜上生长,并具有分裂增殖能力。Aim: The purpose of this study was to evaluate the feasibility of using Chitosan-Collagen/Chondroitin sulfate membrane as corneal biological carrier. Methods: According to the rules of asepsis, the shallow corneal limbus was detached and cut into small pieces which were cultured on plate containing chitosan on the bottom. The morphological feature of the cultured limbal epithelia with light-microscope and electron microscope was studied. The cultured cells was differentiated by immunohistological staining AE1/AE5. Results: At 24～48 h after plantation, the cells moved onto the membrane from the small pieces of tissue, the area of cell membrane gradually expanded, the cells contacted one another closely by apophysis at 7^(th) day. The results of immunohistological staining AE1/AE5 were positive. Conclusion: The membrane is biologically fit with rabbits' corneal epithelia, and is suitable for epithelial growth.

关 键 词：壳聚糖共混膜 兔角膜缘上皮细胞 体外培养 生物学鉴别 

分 类 号：R772.2[医药卫生—眼科]