条件培养基对日本血吸虫卵黄培养细胞Ag-NORs的影响  

Effects of Conditioned Medium on Ag-NORs in Cultured Vitelline Cells from Schistosoma Japonicum

作　　者：罗超[1] 陈喜珪[1] 赵桂花[1] 

机构地区：[1]湖北科技学院基础医学院,湖北咸宁437100

出　　处：《湖北科技学院学报（医学版）》2012年第6期461-464,共4页Journal of Hubei University of Science and Technology(Medical Sciences)

摘　　要：目的以核仁组织区相关嗜银蛋白(Ag-NORs)为评价指标,观察条件培养基对日本血吸虫卵黄培养细胞的影响。方法灌注法获取28d虫龄的日本血吸虫虫体,雌雄分离后冷消化法制备细胞悬液,调细胞为1×106个/ml,联合法接种细胞。将接种于小盖玻片上的血吸虫卵黄细胞随机分为对照组和实验组2组。对照组用常规培养基即RPMI-1640含20%小牛血清附加常量抗生素(青霉素1×105U/L,链霉素100mg/L)培养,实验组培养基为常规培养基与培养的鼻咽癌细胞上清液的条件培养基1∶1的混合液。在培养的第7d、14d、21d、28d、35d时,定时对各组卵黄培养细胞进行Ag-NORs染色,光镜下观察、拍照,同时随机选取50个左右染色细胞输入HPIAS-2000图像分析仪测定Ag-NORs光密度值,并做统计分析。结果培养第1～5周,实验组细胞Ag-NORs均深于对照组细胞;随着培养时间的延长,两组细胞核内着色颗粒均逐渐减少;两组培养细胞的Ag-NORs含量均逐渐降低,对照组下降较快,实验组下降较缓。相同培养时间比较,实验组培养细胞的Ag-NORs含量均显著高于对照组(P<0.05)。实验组细胞之间两两比较,除第21d与28d(P>0.05)无显著性差异外,其余各组差异均显著(P<0.01);对照组细胞之间两两比较,除第28d与35d外(P>0.05),其余各组差异均显著。结论条件培养基不仅可促进卵黄培养细胞的分裂增殖,而且能保持卵黄细胞在培养过程中的分裂增殖活性。Objective To study the effects of conditioned medium on the growth and proliferation of cultured vitelline cells from Schistosoma japonicum.Methods The 28-day schistosoma of S.japonicum were harvested by perfusion,vitelline cells were made by cold digestion,and then inoculated on the small cover glass.The cultured cells were randomly divided into test and control groups.The control group was cultured by the routine medium,the test group was cultured by the conditioned medium.7 days later,the cells were stained by the silver colloid technique to show Ag-NORs from 1 week to 5 weeks.The content of Ag-NORs in the cultured cells was measured by photometer-image analytical instrument.Results The observation of light microscope showed that the staining of nucleus became weaker,the staining granules in the nucleus decreased.The results of photometer-image analysis showed that the content of Ag-NORs was decreased with the extending of culture time.The content of Ag-NORs in the cells of control group decreased sharply than that of test group.At the same culture time,the content of Ag-NORs in test group was stronger than that of control group（P0.05）.Conclusion The conditioned medium can significantly stimulate the proliferation of the cultured vitelline cells.

关 键 词：日本血吸虫 条件培养基 卵黄细胞 核仁组织区相关嗜银蛋白 

分 类 号：R383.24[医药卫生—医学寄生虫学]