皮肤成纤维细胞构建组织工程化猪角膜基质样组织  

Construction of tissue engineered porcine corneal stroma with skin fibroblasts

作　　者：张艳青[1] 张文杰[1] 胡晓洁[1] 周广东[1] 崔磊[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海交通大学医学院第九人民医院整复外科上海市组织工程重点实验室

出　　处：《上海交通大学学报（医学版）》2007年第2期156-160,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点基础研究项目("九七三"项目)(2005CB522702)~~

摘　　要：目的探讨以皮肤成纤维细胞为种子细胞,在猪角膜微环境中构建组织工程化角膜基质组织的可行性。方法分离获取胎猪皮肤成纤维细胞经体外培养、扩增。通过转染绿色荧光蛋白(GFP)基因标记细胞,示踪细胞的转归。选取第3代的细胞接种于可吸收生物材料聚羟基乙酸(PGA),形成细胞-生物材料复合物,体外培养1周后,移植于猪的角膜基质层内,于第8周取材进行大体、组织学检查及超微结构观察。以角膜基质细胞为种子细胞构建的组织工程化角膜基质组织作为对照。结果术后8周,实验组角膜逐渐恢复透明,与对照组角膜无明显差别。组织学检查显示形成新生的角膜基质样组织,呈板层状,排列较规则,与对照组角膜组织相似。超微结构观察及胶原纤维直径检测显示,实验组与对照组角膜组织差异无统计学意义。荧光显微镜观察到GFP阳性细胞存在。结论皮肤成纤维细胞可能替代角膜基质细胞,在猪角膜内构建角膜基质样组织。Objective To explore the feasibility of constructing tissue engineered porcine corneal stroma with skin fibroblasts in vivo. Methods Skin fibroblasts were isolated from embryonic porcine, cultured and expanded in vitro. Cells were labeled with green fluorescence protein （GFP） gene by retro-viral infection. Cells at passage 3 were seeded on polyglyeolie acid （PGA） non-woven fibers to form a cell-scaffold complex. The complexes were then implanted into poreines＇ corneal stroma after culturing in vitro for 1 week. Engineered stroma was observed continuously and harvested after 8 weeks for gross and histological evaluation. PGA with corneal stromal cells was served as control. Results The engineered tissue in the stroma gradually became transparent over a period of 8 weeks, showing no difference with the control group. Histologically, the engineered stromal lamellar was relatively regular and similar to the control. The im- planted cells were confirmed by GFP expression under fluorescent microscope. By transmission electron microscopy examination, no significant difference in the diameter of collagen fiber was observed between the engineered stroma and normal stroma. Conclusion Tissue engineered corneal stroma may be formed with skin fibroblasts in porcine corneal mieroenvironment.

关 键 词：组织工程 角膜基质 皮肤成纤维细胞 

分 类 号：R318.1[医药卫生—生物医学工程]