| 摘要: |
| 为了探讨丝裂酶素C处理后鼠胚成纤维细胞(Mouse embryonic fibroblast,MEF)活力的变化规律,试验用10μg/mL丝裂酶素C处理MEF,研究丝裂酶素C处理时间及其处理后细胞接种量、血清浓度和细胞代次对MEF活力的影响。结果表明,用10μg/mL丝裂酶素C处理2和3 h,MEF活力比处理4 h的稳定,处理2 h的MEF活力与处理3 h的没有显著差异;用10μg/mL丝裂酶素C处理3 h后,MEF以(0.8～1.6)×104/孔接种96孔板,其活力比2.0×104/孔和4.0×104/孔接种的稳定;用含体积分数10%犊牛血清培养液培养的MEF活力,比用含体积分数2%,5%,15%和20%犊牛血清培养液培养的稳定;第1,3代MEF活力比第5,7代的稳定。表明,用10μg/mL丝裂酶素C处理2～3 h,第1,3代MEF以(0.8～1.6)×104/孔接种96孔板,用含体积分数10%犊牛血清培养液培养,是利用丝裂酶素C制备MEF饲养层的适宜条件。 |
| 关键词: 胎鼠成纤维细胞；MTT 丝裂霉素C；细胞活力 |
| DOI： |
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| 基金项目:国家“863”计划项目（2005AA21905）；教育部重大项目（03160）；陕西省重大科技专项（2006KZ05-G1） |
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| Analysis of vital maintenance of mouse embryonic fibroblasts after treatment of mitomycin |
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| Abstract: |
| This paper analyzed cell vitality and maintence time of MEF after treatment of mitomycin C by means of MTT method,thereby compared effects of treating time of mitomycin C,cell plating densities,serum concentration,and cell passages on MEF vitalities after mitomycin C treatment.The results suggested that cell vitality sustained more stablely after 2 hours and 3 hours than 4 hours of mitomycin treatment at the concentration of 10 μg/mL.After inactivation with mitomycin C at the concentration of 10 μg/mL,cell vitality sustained more stablely at the plating densities of (0.8-1.6)×104/well into the 96 well plate than that at 2.0×104/well and 4.0×104/well respectively.Meanwhile,MEF vitality maintained longer cultured in the medium supplemented with 10%（V/V）of newborn cattle serum than 2%,5%,15%,20%（V/V）of newborn cattle serum respectively.Furthermore,MEF vitalities at the first passage and the third passage sustained more stablely than its vitalities at the fifth passage and the seventh passage.At last,with 2-3 hours of mitomycin C at the concentration of 10 μg/mL,MEF at the first passage and the thid passage,plated at the plating densities of (0.8-1.6)×104/well into the 96 well plate in the medium supplemented with 10% new born castle serum was proved to be the fit condition in preparation for MEF feeder layers. |
| Key words: mouse embryonic fibroblast MTT mitomycin C cell vitality |
