人体毛囊角阮细胞培养技术——在3T3细胞饲养层上的原代及传代培养

中华皮肤科杂志 ›› 1989, Vol. 22 ›› Issue (3): 153-155.

人体毛囊角阮细胞培养技术——在3T3细胞饲养层上的原代及传代培养

夏隆庆¹, Lehmann F², Detmar M², Stadler R², Orfanos CE²

1. 中国医学科学院皮肤病研究所;
2. The Department of Dermatology, University Medical Center Steglitz, The Free University of Berlin, Berlin (West), FR Germany

收稿日期:1988-03-10 修回日期:1988-08-01 出版日期:1989-06-15 发布日期:1989-06-15

CULTURE TECHNIQUE FOR HUMAN HAIR FOLLICLE KERATINOCYTES PRIMARY CULTURE AND SUBCULTURES ON 3T3 CELL FEEDER LAYERS

XIA Long-Qing¹, LEHMANN F²

1. Institute of Dermatology, Chinese Academy of Medical Sciences, Nanjing;
2. The Dept of Dermatology, University Medical Center Steglitz, The Free University of Berlin (West), FR Germany

Received:1988-03-10 Revised:1988-08-01 Online:1989-06-15 Published:1989-06-15

摘要/Abstract

摘要： 本文介绍一种人体毛囊外根鞘角阮细胞的培养技术。将拔取的生长期人发放在3T3细胞饲养层上和具有多种生长因子的DMEM中,在37℃,CO₂孵箱中培养。原代培养14天后,先用0.02%EDTA去除3T3细胞。然后,用0.25%胰蛋白酶和0.02%EDTA的棍合液,将培养的角阮细胞消化成分散的单个细胞。再将此细胞悬液放在新制备的3T3细胞饲养层上进行传代培养。通常可传代培养3~4次。作者认为这是一种简便有效的培养方法,可以从原代和传代培养中获得大量人发毛囊角肮细胞,供医学科学研究。

Abstract: A technique for the culture of outer root sheath keratinocytes from plucked human hair follicles has been developed. The anagen-phase human hairs were plated on 3T3 cell feeder layers and were cultured in DAhEM, substituted with different' growth factors in a CO₂ incubator at 37℃. After 14 days in primary culture, 3T3 cells were removed with 0.02% EDTA and the keratinocytes were detached and disaggregated to single cells by treatment with 0.25% trypsin and 0.02% EDTA at 37℃ for 15 min. The dispersed cells were then replated on fresh 3T3 cell feeder layers. The keratinocytes were serially sub-cultured for 3-4 times. By this technique, large amounts of stratified multilayerd kera-tinocyte cultures were readily obtained in primary culture as well as in subcultures. The technique described here is a simple and effestive culture method and provides a large amount of hair follicle keratinocytes for medical research programmes.

夏隆庆, Lehmann F, Detmar M, Stadler R, Orfanos CE. 人体毛囊角阮细胞培养技术——在3T3细胞饲养层上的原代及传代培养[J]. 中华皮肤科杂志, 1989, 22(3): 153-155.

XIA Long-Qing, LEHMANN F. CULTURE TECHNIQUE FOR HUMAN HAIR FOLLICLE KERATINOCYTES PRIMARY CULTURE AND SUBCULTURES ON 3T3 CELL FEEDER LAYERS[J]. Chinese Journal of Dermatology, 1989, 22(3): 153-155.