鼠胚胎成纤维细胞饲养层及与鼠尾胶联合促日本血吸虫培养细胞贴壁作用的研究  

Effects of mouse embryonic fibroblast feeder layers and combined of mouse embryonic fibroblast and collagen of rat tail on adhesion of cultured cells from Schistosoma japonicum

作　　者：李俊琳[1] 熊飞[1] 钟沁萍[1] 明珍平[1] 董惠芬[1] 蒋明森[1] 

机构地区：[1]武汉大学基础医学院人体寄生虫学教研室,血吸虫病研究室,430071

出　　处：《中国地方病学杂志》2006年第2期145-149,共5页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目(30570243);湖北省卫生厅资助项目(JX1B067);武汉大学杰出青年基金资助项目(20030515)

摘　　要：目的观察鼠胚胎成纤维细胞(MEF)与鼠尾胶及联合对日本血吸虫培养细胞的促贴壁作用。方法灌注法获取21 d虫龄的日本血吸虫虫体,冷消化法制备细胞悬液,调细胞数为1×109／L,并将细胞分别接种于预先铺敷鼠尾胶(鼠尾胶组)、MEF(MEF组)、铺敷鼠尾胶后接种MEF(联合组)和既不铺敷鼠尾胶也不接种MEF(对照组) 的4组培养瓶中培养,培养基为RPMI-1640含20％小牛血清附加常量抗生素。定时计数贴壁细胞数,计算贴壁率。结果随着培养时间从24-48 h,各组日本血吸虫细胞的贴壁率逐渐增高,48 h以后贴壁率开始下降;方差分析显示,同组内不同培养时间细胞贴壁率比较,差异有统计学意义(F=149．22、81．97、232．86、63．05,P<0．01);q检验发现,同组内不同培养时间两两比较差异有统计学意义(P<0．01)。相同培养时间,各组间培养细胞的贴壁率不都相同 (F24=44．65,P<0．01;F48=1 071．489,P<0．01;F72=4 208．35,P<0．01);q检验分析显示,除培养24 h的MEF组与鼠尾胶组细胞的贴壁率差异无统计学意义外(q=0．792,P>0．05),其余两两比较均有统计学意义(P<0．01)。结论 MEF对日本血吸虫培养细胞的贴壁具有促进作用,MEF与鼠尾胶联合对培养细胞的贴壁具有协同作用。Objective To study the effects of mouse embryonic fibroblast （MEF） feeder layers and the combination of MEF and collagen of rat tail on adherence of the cultured cells from Schistosomajaponicum （S. japonicum）. Methods The 21 days schistosomula of S. japonicum were harvested by perfusion. Cell suspension was made by cold digestion and diluted to a concentration of 1 × 10^9 cells/L. Then, the cells were inoculated and cultured on the walls of culture flasks that had been previously smeared with collagen of rat tail or/and cultured MEF and were divided into collagen group, MEF group and collagen combined with MEF group, respectively. The cells cultured without collagen of rat tail and MEF was set up as control group. The number of the adherent cells was counted timely, while the rate of cell adherence was calculated. Results It was found that the rate of cell adherence increased gradually along with the extending of the culture times from 24 to 48 hours, but decreased after 48 hours. Analysis of variance showed that the rates of cell adherence had differences in different culture time （F = 149.22, 81.97, 232.86, 63.05, P 〈 0.01 ）. q-test analysis revealed that the rates of cell adherence in the same group had significant differences in different culture time （P 〈 0.01）. At the same culture time, analysis of variance showed that the rates of cell adherence had dhq＇erence among the four groups （F24 = 44.65 ,P 〈 0.01 ;F48 = 1 071.489,P 〈 0.01 ;F72 = 4 208.35 ,P 〈 0.01 ）. q-test analysis revealed that there was a significant difference rate of cell adherence（P 〈 0.01 ） except for that between MEF group and collagen group cultured for 24 h （q = 0.792, P 〉 0.05）. Conclusions MEF has obvious effects on the adherence of cultured cells of S. japonicum, and the combination of MEF and collagen of rat tail has synergetic effects on the adherence of cultured cells.

关 键 词：血吸虫 日本 细胞 培养的 贴壁率 

分 类 号：R383.24[医药卫生—医学寄生虫学] R619.6[医药卫生—基础医学]