不同接种密度对人表皮细胞培养融合的实验研究  

The research of human keratinocytes cultured in vitro at different seeding concentration

作　　者：于大山[1] 冯光珍[1] 

机构地区：[1]武警总医院烧伤整形科,北京市100039

出　　处：《中国美容医学》2003年第2期136-137,T001,共3页Chinese Journal of Aesthetic Medicine

摘　　要：目的:研究不同密度接种对人表皮细胞体外培养融合的影响。方法:外科切取无菌皮肤或包皮环切手术标本,经两步酶消化处理后,制备单个表皮细胞悬液。以不同细胞密度接种于含有3T3滋养细胞的培养瓶中,实验室体外原代培养扩增。结果:观察不同接种密度表皮细胞的体外培养增殖,结果表明人表皮细胞具较高的生长、增殖、融合能力。2×106cell/75cm2培养瓶密度接种人表皮细胞于含3T3滋养细胞培养瓶中,表皮细胞贴壁良好,细胞融合时间较短。结论:鼠3T3细胞能促进人表皮细胞贴壁,仍不失为较好的表皮细胞培养滋养层。适宜的表皮细胞接种密度为2×106cell/75cm2培养瓶。Objective: To investigate the effects of different seeding concentration performed on human keratinocytes.Methods: Sterile skin biopsy or foreskin were treated with DispaseⅡand 0.125%trypsin/0.01%EDTA1:1 mixture to form single keratinocytes susupension.then the cells were seeded into 75cm2flask(containing 3T3 feeder layer)at different concentration and cultured in vitro. Results: Cultured the human keratinocytes in vitro and observe it's proliferation. The results show human keratinocytes have high ability to growth,proliferation and confluence. In primary culture,seeded human keratinocytes in concentration of 2×106 cell/75cm2 flask(containing 3T3 feeder layer),the attachment and growth are good and the time of confluence are shorten.Conclusion: 3T3 cells can accelerate the attachment of human keratinocytes,It is a good feeder layer for human keratinocytes culture. And the proper seeding concentration is 2×106 cell/75cm2 flask in primary culture.

关 键 词：人表皮细胞 细胞培养 3T3细胞 接种密度 

分 类 号：R622.1[医药卫生—整形外科]