不同刺激剂PHA和ConA对绒山羊淋巴细胞有丝分裂中期化的影响  

Effect of Phytohaemagglutinin and Concanavalin A in Different Stimulants on Mitosis Metaphase of Cashmere Goat Lymphocyte

作　　者：俎红丽[1] 张博伦[2] 于梦然 蔡婷[1] 王琳[1] 欧阳效晴[1] 刘志红[1] 李金泉[1] 

机构地区：[1]内蒙古农业大学动物科学学院、动物遗传育种与繁殖重点实验室,内蒙古呼和浩特010018 [2]复旦大学生命科学院,上海200433

出　　处：《中国畜牧兽医》2014年第10期118-122,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家863计划(2013AA102506);国家自然科学基金(31072005、31272421);国家农业部现代农业产业技术体系(CARS-40-05)

摘　　要：试验旨在探索使较高比例的淋巴细胞富集在有丝分裂G2/M期的最佳条件。运用植物血凝素(PHA)和刀豆蛋白A(ConA)对淋巴细胞进行刺激使其增殖,培养一定时间后加秋水仙素对淋巴细胞进行同步化处理,用流式细胞仪检测G2/M期的细胞数量进行比对,观察试剂的最佳作用浓度和加秋水仙素的最佳时间。结果表明,采用PHA和ConA刺激淋巴细胞增殖的最佳作用浓度是60和5μg/mL,且淋巴细胞经ConA刺激培养45h再加秋水仙素处理5hG2期的比例最高为58.38%。结果提示,就绒山羊的淋巴细胞来说,ConA刺激绒山羊淋巴细胞增殖的效果比PHA好,且摸索出细胞G2/M期的时间点至关重要。This experiment was conducted to explore optimized conditions to make more Cashmere goat lymphocyte which had been treated in G2/M.Phytohaemagglutinin（PHA）and concanavalin A（ConA）reagents were used to stimulate the proliferation of lymphocytes,the colchicine reagent were used at specific times of lymphocytes cell cycle synchronization and the number of the cell in G/M was measured by flow cytometry to observe the optimal concentration and action time.The results showed that the PHA and ConA optimal concentration to stimulate the proliferation of lymphocytes were 60 and 5μg/mL.In addition,lymphocytes with ConA stimulation culture 45 h,then colchicines was used to treat 5h.The percentage of lymphocytes in G2/M accounted for 58.38%.The results suggested that the treatment effect of ConA worked better than PHA in proliferation of lymphocytes.

关 键 词：绒山羊 植物血凝素(PHA) 刀豆蛋白A(ConA) 淋巴细胞 秋水仙素 细胞同步化 流式细胞仪 

分 类 号：Q253[生物学—细胞生物学]