摘要
从三带喙库蚊(Culex tritaeniorhynchus)初孵幼虫组织建立了一个细胞系(CT-188),细胞生长于改良199培养基中。蚊卵的表面消毒方法简化为仅以70％酒精消毒10min。组织块接种后10～11d,可形成原代单层。传代细胞接种后,可在30min内贴壁,在48h达到分裂高峰,36～84h为细胞的对数生长期。细胞类型以梭形细胞为主(约占95％),掺杂少数圆形及不规则细胞。细胞核学特征以二倍体(2n=6)为主(约占80％以上)。目前,细胞已传至34代,均冷冻保存于液氮(-196℃)中,且复苏成功。经鉴定本细胞系无病毒及支原体污染。
The establishment and growth of a cell line(CT-188) from fresh incubated larvae of Culex tritaeniorhynchus in a modified 199-medium is described. Only ethanol (70%)were used for sterilizing rather than the complex of mercuric chloride(HgCl_2) and ethanol. The majority of CT-188 cells were a fibroblast-type (approximately 95%); however, a small number of epithelial-like cells and other cells were also present in the culture. 10~11 days after the primary cells had covered the bottom of the container and could be subcultured. After seeding cells began to attach to the glass surface of the container within 30 minutes and reached the division peak in 48 hours. Most of the cells had the normal 2n chromosome complement of 6, although on occasion cells with more this number were seen. The cells are stored in liquid nitrogen(-196℃)at various passage levels and have been successfully resuscitated.
关键词
三带喙库蚊
蚊
细胞系
细胞培养
Culex tritaeniorhynchus
mosquito
cell lin
cell culture
