Establishment of a novel method for primary culture of normal human cervical keratinocytes

Establishment of a novel method for primary culture of normal human cervical keratinocytes

LIU Yu-zhen;L(U) Xiu-ping;PAN Zi-xuan;ZHANG Wei;CHEN Zhao-ri;WANG Hui;LIU Hua

【期刊名称】《中华医学杂志（英文版）》 【年(卷),期】2013(126)017

【摘要】Background Cervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10％ fetal bovine serum (FBS).However,it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth

medium

alone

because

cell

attachment

can

be

poor.Therefore,the culture of these cells is extremely difficult.In this study,we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.Methods Normal cervical epithelial tissue pieces were obtained and digested with type Ⅰ collagenase to dissociate the cells and a single cell suspension produced.The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type Ⅰ from rat tail,with modified keratinocyte serum-free medium (K-SFM) supplemented with 5％ FBS.After attachment,the medium were replaced with K-SFM without FBS.The expression of basal keratins of the ectocervical

epithelium,K5,K14

and

K19

were

assayed

by