Cell-culture system for continuous production of Toxoplasma gondii tachyzoites
Cell-culture system for continuous production of Toxoplasma gondii tachyzoites
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DOI：
10.1007/s100960050423
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发表时间：
1999-12-01
影响因子：
4.5
通讯作者：
Ho-Yen, DO
中科院分区：
文献类型：
--
作者：
Evans, R;Chatterton, JMW;Ho-Yen, DO
本研究的目的是确定一个可持续的细胞系和培养方法，可以连续提供足够数量的弓形虫速殖子，以满足综合医院实验室的需要。研究了三种连续细胞系（HeLa、LLC和Vero）和三种细胞培养方法（常规烧瓶培养、膜基烧瓶培养和自动培养系统）。在多重感染和时间过程实验中，HeLa是选择的细胞系。从HeLa细胞收获的速殖子产量显著高于LLC细胞（P < 0.00005）或Vero细胞（P < 0.05）。基于膜的培养瓶最初的产率（6.15 × 10（6）速殖子/ml）高于常规培养瓶（1-2 × 10（6）速殖子/ml），但这些产率并不持久。自动化细胞培养系统不适合寄生虫培养。在25 cm（2）烧瓶中连续传代是成功的，产生1 x 10（6）速殖子/ml;在38次传代中，感染96-120 h后活力超过90%，在此期间活力提高，收获时间变得更加一致。在HeLa细胞中连续培养的弓形虫可以提供有规律的活速殖子供应。HeLa衍生的速殖子可用于染料试验的证明证实了该体外系统用于综合医院实验室的潜力。
The aim of this study was to identify a sustainable cell line and culture method that could continuously provide a sufficient quantity of Toxoplasma gondii tachyzoites to serve the needs of a general hospital laboratory. Three continuous cell lines (HeLa, LLC and Vero) and three cell-culture methods (culture in conventional flasks, culture in membrane-based flasks and an automated culture system) were investigated. In multiplicity of-infection and time-course experiments, HeLa was the cell line of choice. Harvests from HeLa cells had significantly higher tachyzoite yields than those from LLC cells (P < 0.00005) or Vero cells (P < 0.05). Membrane-based flasks gave higher yields (6.15 x 10(6) tachyzoites/ml) than conventional flasks (1-2 x 10(6) tachyzoites/ml) initially, but these were not sustained. The automated cell-culture system was unsuitable for parasite culture. Continuous passage in 25 cm(2) flasks was successful, yielding 1 x 10(6) tachyzoites/ml; viability exceeded 90% after 96-120 h of infection throughout 38 passes, during which time the viability improved and the time to harvest became more consistent. Toxoplasma gondii grown in continuous culture in HeLa cells can provide a regular supply of viable tachyzoites. Demonstration that HeLa-derived tachyzoites could be used for the dye test confirms the potential of this in vitro system for use in general hospital laboratories.