摘要
目的 建立用包皮成纤维细胞 (Humanforeskinfibroblasts ,HFF)培养弓形虫速殖子的方法。 方法 将包皮环切术切下的包皮在含青、链霉素的Hank’s液中充分洗涤后 ,用眼科剪子剪成 0 .5cm3 的组织小块 ,移入培养瓶中培养 ,待HFF细胞长满瓶底后 ,用胰酶消化并将其接种到培养皿中的盖玻片上培养 ,HFF长满盖玻片后 ,用弓形虫速殖子感染HFF ,镜下观察并于 1、2 .5、4、5 .5、10、2 3和 3 2h取出其中一皿中的盖玻片 ,经姬氏染色、二甲苯透明后 ,镜下观察速殖子生长情况。 结果 弓形虫速殖子大多在感染后 3h～ 5h侵入HFF ,3 2h左右 ,假包囊破裂 ,释放出虫体。 结论 成功建立了用HFF培养弓形虫的方法。
Objective To establish the method of cultivating Toxoplasma gondii tachyzoites in human foreskin fibroblasts(HFF).Methods After washed twice in Hank's containing penicillin (500 U/ml), and streptomycin (500 μg/ml) for 5～10 min each time, the foreskin samples were minced into 0.5 cm 3 tissue pieces, putted into the culture flasks and cultured in modified Eagle's medium supplemented with 10% of fetal bovine serum for 28 days. Before infection, HFF were allowed to adhere to glass cover slips in dishes. Confluent monolayers on glass cover slips were infected with 10 3 parasites in DMEM containing 1%～3% fetal bovine serum. Cultures were examined at frequent intervals with an inverted microscope to document the infection of cells and the formation of pseudocysts. After infection the glass cover slips were taken out from the dishes at 1, 2.5, 4, 5.5, 10, 23, 32 hours respectively and stained with Giemsa . Then photomicrographs were made. Results About 3 h～5 h after infection tachyzoites invade into HFFs. At 32 h after the addition of tachyzoites, pseudocysts were observed in the medium and begin to rupture. Conclusion The method of cultivating T. gondii tachyzoites was successfully established.
出处
《中国寄生虫病防治杂志》
CSCD
2004年第5期268-269,F002,共3页
Chinese Journal of Parasitic Disease Control
关键词
弓形虫
包皮成纤维细胞
细胞培养
Toxoplasma gondii
human foreskin fibroblasts
cell cultivation
