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二甲基苯蒽诱导山羊乳腺上皮细胞体外转化研究
王 莉1, 安志兴1, 张 涌1
西北农林科技大学 生物工程研究所,农业部家畜生殖内分泌与胚胎工程重点开放实验室
摘要:
分别用5,10和15 μg/mL二甲基苯蒽(DMBA)的完全培养液处理体外培养的山羊乳腺上皮细胞36, 24和12 h,再以25 ng/mL佛波酯(TPA)作用2周,研究DMBA和TPA对山羊乳腺上皮细胞的诱导转化作用,确定最佳诱导转化条件,探讨二甲基亚砜(DMSO)对细胞增殖的影响。结果表明,用含15 μg/mL DMBA的完全培养液培养12 h,再用TPA连续作用4 d,几乎无山羊乳腺上皮细胞存活,10 μg/mL DMBA培养24 h转化效果较差,5 μg/mL DMBA培养36 h转化效果最佳;转化的山羊乳腺上皮细胞体积增大、核浓缩、生长排列紊乱、无规则并出现转化灶;DMSO对细胞生长有抑制作用.随培养时间延长抑制作用减弱。试验初步建立了乳腺上皮细胞的体外转化系统。
关键词:  二甲基苯蒽  细胞转化  山羊  乳腺上皮细胞
DOI:
分类号:
基金项目:国家“863”计划项目(2001AA213081)
Transformed study on goat mammary epithelial cells induced by DMBA in vitro
Abstract:
Exposed the goat mammary cells from enzyme digestion to D-MEM/F12 containing 5,10 and 15 μg/mL DMBA for 36,24 and 12 h respectively, then the cells were cultured in D MEM/F12 containing 25 ng/mL TPA for two weeks,studied the result of goat mammary epithelial cells induced by DMBA and TPA,ascertained the best induced condition,and discussed the cell proliferous influence by DMSO.The result shows,the group cultured by 15 μg/mL DMBA after 12 h was almost dead;the group treated with 10 μg/mL DMBA after 24 h showed the worse effect of translation;the group treated by 5 μg/mLDMBA after 36 h showed the better effect of translation.The characters of translated cells were augmenting and flat,then the karyons of cell were concentrating,the size of cell were increasing at the same time,the array of cell were disorganized and the focus of translation appeared;the restraining effect of DMSO reduced as the culture time prolongs.After culturing forty passage,the translatic models of mammary epithelial cell induced by DMBA were established.
Key words:  DMBA,cell translation,goat,mammary epithelial cells

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