人巨噬细胞和人包皮成纤维细胞体外培养弓形虫速殖子  

In vitro culture of Toxoplasma gondii tachyzoites in human macrophage and human foreskin fibroblasts

作　　者：兰兰[1,2] 吴翡斐[2] 梁韶晖[2] 

机构地区：[1]宁波卫生职业技术学院,宁波315040 [2]温州医科大学寄生虫学教研室,温州325035

出　　处：《国际医学寄生虫病杂志》2014年第4期227-229,233,共4页International JOurnal of Medical Parasitic Diseases

基　　金：浙江省自然科学基金（Y2100971）

摘　　要：目的 比较弓形虫RH株速殖子在人巨噬细胞和人包皮成纤维细胞内的增殖情况. 方法 按照弓形虫速殖子∶细胞为1∶1的比例将弓形虫RH株速殖子分别与人巨噬细胞和人包皮成纤维细胞共培养0、6、12、24、48 h后,于显微镜下观察细胞内虫体增殖情况.在两种细胞培养瓶中分别接种弓形虫RH速殖子,连续传代,每隔10代取弓形虫速殖子,将其密度调整为1×10^7个/ml,0.3 ml/只,腹腔接种ICR健康小鼠,记录小鼠存活时间. 结果 在相同条件下弓形虫速殖子侵入人巨噬细胞的时间早于侵入人包皮成纤维细胞的时间;共培养48 h时,绝大部分弓形虫速殖子胀破细胞,游离在细胞外.在两种细胞培养瓶中分别接种3×106弓形虫RH速殖子,约72 h后均可获得大量游离的弓形虫速殖子,2～3d传1代.人巨噬细胞内连续传代约30次,每代可获得（1×10^7～4×10^7）个速殖子,约增殖3～13倍;人包皮成纤维细胞中连续传代30次,每代可获得（1×10^7～3×10^7）个速殖子,约增殖3～10倍.弓形虫速殖子传代次数为10、20和30代时,人巨噬细胞内传代的弓形虫速殖子接种小鼠的存活时间分别为（4.3±0.5）、（4.0±0.8）、（4.3±1.2）d,人包皮成纤维细胞内传代的弓形虫速殖子接种小鼠的存活时间分别为（4.0±0.0）、（3.7±0.9）、（4.3±0.5）d,各代之间差异无统计学意义（P＞0.05）,弓形虫速殖子毒力未见明显改变. 结论 弓形虫RH株速殖子可在人巨噬细胞和人包皮成纤维细胞内增殖并长期稳定传代.Objective To compare the proliferation of Toxoplasma gondii RH strain tachyzoites in human macrophage and human foreskin fibroblasts.Methods Toxoplasma gondii RH strain tachyzoites were inoculated into human macrophage and human foreskin fibroblasts according to the rate of 1 ∶ 1,the proliferation of tachyzoites was observed under microscope at 0,6,12,24,48 hour after inoculation.At the same time,the long-term in vitro culture of Toxoplasma gondii tachyzoites maintained in human macrophage and human foreskin fibroblasts was established.The Toxoplasma gondii RH strain tachyzoites were taken out at every 10 generations then inoculated intraperitoneally into ICR mice according to 3 ×10^6 tachyzoites per mice.Then the survival days of the mice were observed.Results The time of Toxoplasma gondii tachyzoites invading into human macrophage is earlier than that into human foreskin fibroblasts.At 48 hour after co-culture,most of the two cells were destroyed by tachyzoites.In the long-term culture system,a large amount of free tachyzoites could be obtained after 72 h incubation.In human macrophage,（1×10^7～4×10^7 tachyzoites could be obtained each generation for 30 generations in continually culture with the proliferation rate of 3-13 times while 1 ×10^7～3×10^7 tachyzoites obtained each generation for 30 generation in HFF cells with the proliferation rate of 3-10 times.So the proliferation of tachyzoites was stable in both cells.The survival time of the mice inoculated with tachyzoites at 10,20 and 30 generation cultured in human macrophages was （4.3＋ 0.5） d,（4.0＋0.8） d and （4.3＋1.2） d respectively,while it was （4.0＋0.0） d,（3.7＋0.9） d and （4.3＋0.5） d respectively for the mice infected with tachyzoites cultured in HFF cells.The difference among various generations was not significant statistically （P〉0.05） and their virulence to mice showed no decrease（P〉0.05）.Conclusion The proliferation of Toxoplasma gondii RH strain tachyzoites cultured in human macrophage and h

关 键 词：弓形虫 速殖子 人巨噬细胞 人包皮成纤维细胞 体外培养 

分 类 号：R382[医药卫生—医学寄生虫学]