【作者】 徐征；

【导师】 罗祺；

【作者基本信息】 吉林大学 ， 外科学， 2008， 硕士

【摘要】 目的:观察成纤维细胞(Fibroblast,FB)复合在壳聚糖/甘油磷酸钠凝胶(chitosan and glycerophosphate,C/GP)表面后,成纤维细胞的生长状态,评估壳聚糖/甘油磷酸钠凝胶作为组织工程支架的可行性。方法:采用胶原酶消化法分离兔FB,然后培养纯化,绘制细胞的生长曲线。并通过倒置显微镜、光学显微镜对成纤维细胞在纤维蛋白胶表面的生长情况进行形态学观察。免疫组化方法检测细胞波形蛋白的表达,鉴定细胞。采用SPSS12.0软件包对数据进行统计学分析。结果:兔成纤维细胞在壳聚糖/甘油磷酸钠凝胶表面铺展形态佳,增殖速度快。细胞生长2-4h贴壁,3d时细胞呈漩涡状,基本完全融合。约5d后细胞逐渐呈复层生长,具有典型的成纤维样细胞外观。免疫组化波形蛋白阳性,差异有显著性(P<0.05),证实细胞来源可靠。结论:由成年兔皮肤可分离培养出足量的原代成纤维细胞;兔成纤维细胞在壳聚糖/甘油磷酸钠凝胶表面生长、增殖良好,壳聚糖/甘油磷酸钠凝胶具有良好的组织相容性,有望成为成纤维细胞生长的支架材料。更多还原

【Abstract】 Objective: To observe the growing state of fibroblasts which were seeded on the chitosan and glycerophosphate(C/GP) gel and evaluate the feasibility of the thermosetting C/GP gel as tissue engineering scaffold.Methods:Adult rabbit skin fibroblast were isolated, cultured and identified in vitro. The rabbit FB were obtained by digesting collagen hypoderm and purified by differential speed attachment. Draw the growth carve of FB of the third generation. Identify FB by vimentin immunostaing.The gel was based on C/GP mixed in 7:1 volume ratio. Cellular morphologies of fibroblasts which were seeded on the C/GP gel were observed under invert microscope and optical microscope, and the cells were identified by vimentin immunostaining analysis. The data were analyzed using SPSS12.0 software package.Results: FB has good expanding form and proliferating trend. The cells appear the typical FB appearance. The growth carve appears the typical“S”. The results of vimentin immunostaing approve the culture cells are rabbit FB.After the combination of FB and C/GP, adult rabbit fibroblasts expanded well and proliferated soon on the C/GP gel. The cells grew adherently on the C/GP in about 2 to 4 hours, at the 3rd day most of the cells converged into ieces, presenting whirlpool shape. After 5 days, duplicate layers cells gradually formed.The cells were positively stained for vimentin. The difference was significant (P<0.05).Conclusions: The primary adult rabbit fibroblasts cells could be obtained by isolation and culture of the adult rabbit skin tissue.Rabbit fibroblasts had a good quality of adherency, transference, multiplication and differentiation on the surface and in the vivo of C/GP materia. C/GP gel may be a suitable material as a scaffold compounded with fibroblasts.更多还原