用HaCaT细胞和正常人黑素细胞构建组织工程皮肤  

Reconstruction of tissue engineered skin with HaCaT cells and melanocytes in vitro

作　　者：曹玉萍[1] 周武庆[1] 马鹏程[1] 刘维达[1] 李玲珺[1] 陶玥 桑红桂[1] 李阿梅[1] 邵雪宝[1] 孙建方[1] 

机构地区：[1]中国医学科学研究院北京协和医学院皮肤病研究所,江苏南京210042

出　　处：《毒理学杂志》2009年第5期341-344,共4页Journal of Toxicology

基　　金：江苏省自然科学基金资助项目(BK2008092)

摘　　要：目的探索用人永生化表皮细胞（HaCaT）和黑素细胞复合鼠尾胶原凝胶支架构建组织工程皮肤的可行性。方法HaCaT细胞和黑素细胞与鼠尾胶原复合,经历液下和气液面共13 d的培养,苏木素-伊红（HE）染色观察不同培养阶段组织学形态变化,S-100免疫组化染色观察黑素细胞分布情况。结果随培养时间的延长,组织工程皮肤的细胞层数逐渐增加,结束时可达12-15层;同时,细胞层内发生黑素化,颜色逐渐加深,组织学结构可见黑素颗粒分布广泛,S-100免疫组化染色显示黑素细胞均匀分布于基底层。结论在鼠尾胶原凝胶支架表面用HaCaT细胞和黑素细胞可构建出组织工程皮肤,为探索基于组织工程皮肤的化合物光毒性检测奠定基础。Objective To reconstruct the tissue engineered skin with HaCaT cells and melanocytes on rat tail tendon collagen gel support in vitro.Methods HaCaT cells and melanocytes were combined to rat tail tendon collagen,which then were cultured in the medium for 3 d and at the air-liquid interface for 10 d.Morphology and histology of the tissue engineered skin during different culture stages were examined with HE staining,and immuno-histo-chemical staining for S100 was conducted to analyze the distribution of melanocytes.Results As time goes by,the cell layer became thicker and thicker,and could reach 12～15 cell layers till the end of culture;Moreover,the tissue engineered skin was pigmented and the tanning degree was getting stronger.The melanins were distributed widely in histology and S-100 positive cells,the melanocytes,were distributed uniformly in the basal layer.Conclusion Reconstruction of the tissue engineered skin with HaCaT cells and melanocytes on rat tail tendon collagen gel support in vitro was successful,which was useful for studying the phototoxicity testing based on tissue engineered skin.

关 键 词：HACAT细胞 黑素细胞 组织工程皮肤 光毒性 

分 类 号：R318.19[医药卫生—生物医学工程]