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小鼠心肌组织裂解上清液诱导小鼠骨髓瘤Sp2/0细胞凋亡

时间:2018-01-20 药学毕业论文 我要投稿
作者:王玮,张文艳,吴建军,类延花,金晶,方草晖,胡勇,王明丽
【关键词】 心肌上清液
Apoptosis of mouse myeloma Sp2/0 cells induced by supernatant of mouse cardiac muscle lysate
  【Abstract】 AIM: To study the roles of mouse cardiac muscle lysate supernatant in inducing the apoptosis of mouse myeloma Sp2/0 cells. METHODS: Various concentrations (1∶6, 1∶12, 1∶24) of mouse cardiac muscle lysate supernatant and cyclophosphamide were added into the Sp2/0 cell culture wells. After 24 h, microscope was used to observe the morphological changes of Sp2/0 cells and apoptosis rate was determined by flow cytometry after 48 h. Morphological changes of Sp2/0 cells cocultured with mouse cardiac muscle cells were observed after 10 h. The apoptosis of Sp2/0, Hela, Hep2 and Vero cells was induced with various concentrations (1∶5, 1∶10, 1∶20 and 1∶40) of mouse cardiac muscle lysate supernatant, liver lysate supernatant, thymus lysate supernatant and cyclophosphamide, and then the difference of apoptotic rate among them was compared. RESULTS: Mouse cardiac muscle lysate supernatant could induce the apoptosis of the Sp2/0 cells and the apoptotic rate increased in a concentrationdependent manner, but there was no difference between mouse cardiac muscle lysate supernatant group and cyclophosphamide group. After a 10hour coculture with mouse cardiac muscle cell lysate, the apoptosis of a few Sp2/0 cells was seen. The mouse cardiac muscle lysate supernatant, liver lysate supernatant and thymus lysate supernatant could all induce apoptosis of the Sp2/0, HeLa, Hep2 and Vero cells. But the effect of the mouse cardiac muscle lysate supernatant was more obvious and permanent. CONCLUSION: The mouse cardiac muscle lysate supernatant can significantly induce apoptosis of mouse myeloma Sp2/0 cells.
  【Keywords】 cardiac muscle lysate supernatant; apoptosis; mouse myeloma Sp2/0 cell
  【摘要】 目的: 研究小鼠心肌上清液诱导小鼠骨髓瘤Sp2/0细胞凋亡的作用. 方法: 在Sp2/0细胞培养孔中,分别加入不同稀释度(1∶6, 1∶12, 1∶24)的小鼠心肌上清液和环磷酰胺, 24 h后镜下观察细胞形态,并于48 h后流式细胞仪检测凋亡细胞的比率;同时,Sp2/0细胞和小鼠心肌细胞共培养,10 h后观察Sp2/0细胞形态学变化;并用不同稀释度(1∶5, 1∶10, 1∶20, 1∶40)小鼠心肌裂解上清液、肝脏裂解上清液、胸腺裂解上清液及环磷酰胺诱导Sp2/0, HeLa, Hep2和Vero细胞凋亡,并比较其差异. 结果: 不同稀释度的心肌裂解上清液均可诱导Sp2/0细胞的凋亡,尤以高稀释度更明显;小鼠心肌细胞与Sp2/0细胞共培养10 h后,少部分Sp2/0细胞出现凋亡;小鼠心肌裂解上清液、肝脏裂解上清液、胸腺裂解上清液均可引起Sp2/0, HeLa, Hep2和Vero细胞发生凋亡,但小鼠心肌裂解上清液作用明显,且作用时间持久. 结论: 小鼠心肌组织裂解上清液具有明显诱导小鼠骨髓瘤Sp2/0细胞凋亡的作用.

  【关键词】 心肌上清液;细胞凋亡;小鼠骨髓瘤Sp2/0细胞
  0引言
  细胞凋亡与肿瘤的发生发展密切相关,已成为研究热点[1],许多预防治疗肿瘤的药物、细胞因子等抑制肿瘤细胞生长的机制之一是诱导肿瘤细胞凋亡. 我们从细胞凋亡的角度探讨心肌上清液诱导小鼠骨髓瘤Sp2/0细胞凋亡.
  1材料和方法
  1.1材料
  Balb/c 20只8周龄小鼠,雌雄各半,体质量约250 g,由安徽安科生物工程股份有限公司动物室提供;环磷酰胺(2 g/L)由湖北科利药业股份有限公司生产; DMEM液由美国生命技术公司生产;胎牛血清由杭州四季青公司提供;流式细胞仪(日本产);Leica荧光显微镜(德国Leica公司);CO2温箱(美国产);紫外分光光度计(上海产). Sp2/0, HeLa, Hep2和Vero细胞株均来源于中国预防科学院病毒所,均以100 mL/L胎牛血清+DMEM液为培养液,按本室常规方法在CO2温箱中培养;乳鼠心肌细胞培养: 取Balb/c新生红皮乳鼠20只,在无菌操作下取出心脏,用PBS冲洗3遍,以眼科剪剪成1 mm3碎块,洗涤,后加2.5 g/L胰蛋白酶液37℃消化20 min,吹打3次,吸取上层细胞悬液,以同样的方法消化3次,直到组织块完全消化为止,以1500 r/m
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