正常人表皮细胞老化过程中的生物学特性(英文)  

Biological characteristics of the normal human keratinocytes during the aging process

作　　者：高学军[1] 蔡霞[2] 张鹏[2] 唐胜建[2] 

机构地区：[1]潍坊医学院附属医院普外科,山东省潍坊市261041 [2]潍坊医学院整形外科研究所,山东省潍坊市261042

出　　处：《中国临床康复》2005年第42期154-156,i0004,共4页Chinese Journal of Clinical Rehabilitation

基　　金：山东省教育厅重点资助项目(03K07)~~

摘　　要：背景:目前有关表皮细胞体外衰老的文献报道甚少,而表皮细胞是构建组织工程化皮肤所必需的种子细胞,文章拟对正常人表皮细胞老化过程中的生物学特性加以阐述,为组织工程化皮肤选择合适的种子细胞奠定基础。目的:通过研究表皮细胞体外增殖与老化规律,为选择合适的组织工程化皮肤种子细胞提供依据。设计:自身对照实验。单位:潍坊医学院整形外科研究所,潍坊医学院附属医院普外科。材料:实验于2000-09/2002-09在潍坊医学院整形外科研究所完成。实验样本来源于在潍坊医学院附属医院普外科行包皮环切术的6～8岁正常男性儿童20例切除的健康包皮组织,监护人获完全同意,将包皮标本用于表皮细胞增殖培养实验。方法:取正常年轻人表皮细胞进行传代培养,以不同代龄细胞为实验对象,采用形态学观察、群体倍增时间、免疫细胞化学及β-半乳糖苷酶染色的方法,检测表皮细胞老化规律。主要观察指标:①表皮细胞生长特性的改变。②表皮细胞形态学改变。③表皮细胞表型的改变。结果:①表皮细胞生长特性的改变:体外单层培养9代,P2的群体倍增时间最短,前5代增殖能力较强,P5以后群体倍增时间明显延长,P8细胞不再增殖。②表皮细胞形态学改变:原代细胞接种3d后开始增殖,4d后细胞加速增殖,培养1周左右细胞接近融合。显微镜、免疫组化鉴定,符合表皮细胞的特点。③表皮细胞表型的改变:随着细胞的连续传代培养,β-半乳糖苷酶组化表达呈现从弱(在年轻细胞中占9%)到强(在老化细胞中占65%)的趋势,β-半乳糖苷酶染色的阳性率和细胞代龄之间呈显著正相关(r=0.87,P<0.01)。结论:①与年轻细胞相比,老化细胞具有衰老形态和酶细胞化学特征的增多;在细胞从年轻向老化发展的进程中,细胞群体倍增时间呈延长的趋势。②与年轻细胞相比,老化细胞中β-半乳糖苷�BACKGROUND：At present, there is still lack of related reports about the aging process of in vitro cultured epidermal cells, since epidermal cells are seed cells necessary for the construction of tissue engineered skin, this articleis is aimed to investigate the biological property of normal human epidermal cells during aging process so as to provide a foundation for the selection of seed cells for tissue engineered skin OBJECTIVE： To observe the in vitro proliferation and aging property of human epidermal cells in order to provide a foundation for the proper selection of seed cells for tissue engineered skin.DESIGN： A self-comparative experiment.SETTING： Orthopedic Surgery Research Instioute of Weifang Medical College and the General Surgery Department of Weifang Medical College Affiliated Hospital.MATERIALS： This experiment was carried out at the Orthopedic Surgery Research Institute of Weifang Medical College, between September 2000and September 2002. Healthy foreskin tissue was obtained from 20 normal boys of 6-8 years old who received peritomy at the General Surgery Department of Weifang Medical College Affiliated Hospital.METHODS： Epidermal cells were obtained from normal young people for subculture. Cells were collected from different culture passages and taken as subjects, and their aging characteristics were assessed through morphological observation, population doubling time （PDT）, immune cytochemistry and beta-galactosidase staining. MAIN OUTCOME MEASURES： ① The changes of the epidermal cell growth characteristics. ② The morphological changes of the epidermal cells. ③ The epidermal cell phenotypic changes. RESULTS： ① The clanges of the epidermal cell growth characteristics： Cells were in vitro cultured by monolayer for 9 passages, and PDT of P2 was the shortest. The cells showed strong proliferation in the first 5 passages.From P6, PDT was obviously prolonged, but the cells from P8 did not proliferate any longer. ② The morphological changes of epidermal cells： The prima

关 键 词：组织工程 表皮/细胞学 细胞 培养的 细胞衰老 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]