人皮肤角质形成细胞的分离与原代无血清培育.pdfVIP

湖南医科大学学报 　2003 ,28 (5) BULL HUNAN M ED UN IV 481 ① 人皮肤角质形成细胞的分离与原代无血清培养 陈 　勇 , 戴和平 , 龙志高 , 文 　娟 , 潘 　乾 , 陈玉祥 , 夏 　昆, 夏家辉 ( 中南大学中国医学遗传学国家重点实验室 ,长沙 4 10078) [摘要] 　目的:建立一种体外人皮肤角质形成细胞分离与原代培养的方法 。方法 :采用两步消化法对皮肤进 行消化 ,获取角质形成细胞进行体外无血清培养基培养 ,进行细胞形态学观察 、免疫组织化学 、透射电镜鉴定及生 长曲线的绘制与分析 。结果 :该方法可获取较多高纯度的角质形成细胞 ,且在体外可快速稳定增殖 。结论 :两步消 化法和体外无血清培养是一种理想的皮肤角质形成细胞分离培养的方法 。 [ 关键词] 　皮肤 ; 　角质形成细胞 ; 　分离 ; 　原代培养 ; 　无血清 [ 中图分类号] 　R322 . 99 1 ; Q343 . 6 　　[文献标识码] 　A 　　　[文章编号] (2003) Isolation and serumf ree culture of human skin primary keratinocytes CHEN Yong , DA I Heping , L ON G Zhigao , et al . ( N ational L aboratory of M edical Genetics , Cent ral S out h University , Changs ha 4 10078 , Chi na) Abstract : 　Objective 　To est ablish an optimal met hod for i n v it ro cult ure and amplification of p rimary keratinocytes from t he human skin . Methods 　The keratinocytes were isolated from t he hu man skin by twostep combined dissociation . The cult ured cells in serumfree media were indentified by morp hology , immunohistochemist ry ,and t ransmissive elect ronmicro scope . Growt h curves were plotted and analyzed . Results 　A lot of highly p ure keratinocytes could be obt ained by t his p rocedure and cul t ured i n v it ro steadily and rapidly . Conclusion 　Twostep combined dissociation and serumfree cul t ure i n v it ro for human skin p rimary keratinocytes is an ideal met hod . Key words : 　skin ; 　keratinocyte ; 　isolation ; 　p rimary cult ure ; 　serumfree [Bull Hunan Med U niv ,2003 ,28 (5) :048104 ] 　　表皮细胞体外培养技术的建立和发展 ,为烧伤 进行实验研究和