用于组织工程构建的表皮角质形成细胞的生物学特性  

Biological Characteristics of Human Keratinocytes Cultured in Tissue Engineering

作　　者：杨光辉[1] 刘伟[1] 崔磊[1] 曹谊林[1] 

机构地区：[1]上海组织工程研究与开发中心

出　　处：《生物医学工程与临床》2003年第2期65-67,F001,共4页Biomedical Engineering and Clinical Medicine

基　　金：国家重点基础研究规划项目 (973项目 ):组织工程的基本科学问题;项目编号G1 9990 5430 4

摘　　要：目的　研究体外培养的人表皮角质形成细胞的生物学特性 ,为构建组织工程皮肤提供技术参数。方法　用Dispase/胰蛋白酶法消化分离人表皮角质形成细胞 ,进行原代及传代培养。计算原代细胞获得率 ,观察各代细胞形态学改变 ,描记细胞生长曲线 ,计算群体倍增时间 (PDT) ,免疫组织化学方法检测细胞角蛋白表达。结果　原代表皮角质形成细胞获得率为 ( 0 .72 6± 0 .3 48)× 10 6 /cm2 ,表皮角质形成细胞体外可培养至第 7代 ,第 2代的PDT最短 ,为( 46.5 7± 2 .2 5 )h ,第 4代的细胞角蛋白表达仍为阳性。结论　综合细胞增殖速率、细胞数量及细胞功能的维持各因素 ,考虑第 3。Objective To study the biological characteristics of in vitro cultured human keratinocytes for providing the technical parameters in constructing tissue engineering skin. Methods The keratinocytes were obtained from human foreskin and isolated by digestion with dispase/trypsin method and then primary and secondary culture were performed. The histological changes of different generations, cell growth curve, population doubling time(PDT) were studied and the expression of cytokeratin was measured with immunochemical method. Results The yield rate of primary keratinocytes was (0.726±0.348)×10 6/cm 2. The in vitro secondary culture of keratinocytes could be succeeded to passage 7. The PDT of passage 2 was the shortest (46.57±2.25) h. The expression of cytokeratin of passage 4 keratinocytes were still positive. Conclusions It is demonstrated that considering of the factors such as cell proliferation rate, cell quantity and function, the passage 3 and 4 of keratinocytes cultured in vitro is the optimal seeding cells for construction of tissue engineering skin.

关 键 词：角质形成细胞 细胞培养 组织工程 生物学特性 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]