人工表皮构建及其结构的研究  

Study on construction of artificial epidermis and research on its structure

作　　者：刘杰[1] 吴小红[1] 崔雪梅[1] 王德文[1] 谷庆阳[1] 张燕[1] 徐龙河[2] 杨颖颙[3] 于淑贤[3] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]解放军总医院,北京100853 [3]中国皮革和制鞋工业研究院,北京100016

出　　处：《军事医学科学院院刊》2002年第4期267-269,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：军事医学科学院创新基金资助项目 (990 5 10 6)

摘　　要：目的 :以角朊细胞作为种子细胞 ,构建人工表皮 ,检测培养的人工表皮的结构 ,为进一步进行动物移植试验奠定基础。方法 :(1)采用分散酶 (dispase)分离表皮细胞 ,以无血清培养法培养表皮细胞。 (2 )人工表皮构建成功后 ,分别进行HE染色及透射电镜观察。 (3)角蛋白免疫组织化学染色。结果 :经过 2～ 3周的培养 ,表皮细胞连接成片 ,呈现典型的“铺路石”状 ;继续培养 5～ 7d后 ,将细胞膜消化下来 ,HE染色显示由 2～ 4层细胞组成 ,电镜可以观察到桥粒及张力微丝束的结构 ;免疫组织化学染色显示培养细胞和细胞膜角蛋白染色阳性。结论 :该人工构建表皮与人正常表皮结构类似 。Objective:To use the keratinocytes as the seeding cells for the tissue engineering of skin in order to make the artificial epidermis, then to examine its structure for the purpose of the further graft experiment.Methods:(1)The dispase was used to isolate the epidermic cells which were then cultured in the serum_free medium.(2)After the epidermis was formed, it was examined by HE staining and transmission electron microscope.(3)Keratin expression was examined by immunocytochemistry.Results:The epidermic cells could grow confluent after 2 to 3 weeks in the serum_free medium, and were like the slabstone.When the epidermic cells were confluent, they were cultured further for 5 to 7 days, and then the skin was made up of 2 to 4 layers.Electronmicroscopic examination found the desmosomes and tonafilament in the skin.Immunocytochemical examination demonstrated the expression of keratin in the cultured cells.Conclusions:The artificial skin is similar to the normal epidermis in the structure, and it can be used as the graft in the further experiments.

关 键 词：人工表皮 皮肤组织工程 皮肤移植 角朊细胞 角蛋白 

分 类 号：R318.1[医药卫生—生物医学工程]