5-aza-dc论文..docVIP

5-aza-dc对延边奶山羊耳成纤维细胞 生物特性的影响 蔡丽娟，尹 多，庄利利，方南洙，李钟淑*、 （延边大学 农学院 动物遗传育种与繁殖实验室，延吉，133002） 摘 要：（目的）探讨DNA甲基化酶抑制剂5-aza-dc对延边奶山羊耳成纤维细胞形态、活率以及染色体的影响，以获得不影响细胞正常生物学特性的最适条件。（方法）5-aza-dc对供体细胞进行不同处理后，利用MTT选定5-aza-dc的最适作用浓度和时间，应用PI、Hochest33342双染和琼脂糖凝胶电泳检测细胞凋亡情况，通过核型分析检测5-aza-dc对细胞染色体的影响。（结果）结果表明，低浓度的5-aza-dc（≤0.01μmol/L）作用72h对细胞影响不大；随其浓度的增加，细胞形态、活率、凋亡及染色体都受到显著影响。（结论）5-aza-dc对细胞的最佳处理时间为72h，此时低浓度的5-aza-dc并未引起细胞凋亡、核型发生改变。 关键词：5-aza-dc；延边奶山羊耳成纤维细胞；细胞活率；细胞形态；细胞凋亡；染色体 中图分类号：S827 文献标识码：A 文章编号： Effect of the cell biological characteristics of 5-aza-dc on Yanbian dairy goat ear fibroblast cell CAI Li-Juan, YIN Duo, ZHUANG Li-Li, FNAG Nan-Zhu, LI Zhong-Shu* (Animal genetic breeding lab, YanBian University, YanJi, China, 133002) Abstract: (Objective) Discuss the effect of DNA methylation inhibitor 5-aza-dc on cell morphology, viability chromosomal of Yanbian dairy goat ear fibroblast cells . Then attempt to obtain an optimal conditions that does not affect the normal cell biological characteristics. (Method) After different treatments of donor cells with 5-aza-dc, used MTT to select the most suitable treatment concentration and time, PI and Hochest33342 double staining, agarose gel electrophoresis experiment were used to detect apoptosis of the cells, applied karyotype analysis to measure the effect of 5-aza-dc on the chromosomes . (Result)) The results showed， low concentrations (≤0.01 μ mol/L ) of 5-aza-dC with 72 h had little impact on the cells; With the increasing of concentration, cell morphology, viability, apoptosis and chromosomes were significantly affected . (Conclusion) The best processing time of 5-aza-dc was 72h. low concentration of 5-aza-dc did not lead to apoptosis and the change of karyotype. Key words: 5-aza-dc; Yanbian dairy goat ear fibroblast cell; cell morphology; cell viability; cell apoptosis; chromosomal 体细胞核移植，是哺乳动物胚胎工程的主要组成部分，也一直是研究的热点。然而核移植研究中仍然存在这许多问题，如核移植效率低、胚胎发育异常、流产、畸形等。大量研究表明，体细胞核移植所用的供体细胞（成纤维细胞、颗粒细胞等）都是高度分化的细胞，具有较高的甲基化水平，在与受体细胞融合时需要经过充