Evans R, Chatterton J M, Ashburn D, Joss A W, Ho-Yen D O
Microbiology Department, Raigmore Hospital, Highland Acute Hospitals NHS Trust, Scotland, UK.
Eur J Clin Microbiol Infect Dis. 1999 Dec;18(12):879-84. doi: 10.1007/s100960050423.
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 (6.15x10(6) tachyzoites/ml) than conventional flasks (1-2x10(6) tachyzoites/ml) initially, but these were not sustained. The automated cell-culture system was unsuitable for parasite culture. Continuous passage in 25 cm2 flasks was successful, yielding 1x10(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.
本研究的目的是确定一种可持续的细胞系和培养方法，能够持续提供足够数量的刚地弓形虫速殖子，以满足综合医院实验室的需求。研究了三种连续细胞系（HeLa、LLC和Vero）和三种细胞培养方法（常规培养瓶培养、基于膜的培养瓶培养和自动化培养系统）。在感染复数和时间进程实验中，HeLa是首选的细胞系。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量（6.15×10⁶个速殖子/毫升）高于常规培养瓶（1-2×10⁶个速殖子/毫升），但产量未能持续。自动化细胞培养系统不适用于寄生虫培养。在25平方厘米培养瓶中连续传代成功，产量为1×10⁶个速殖子/毫升；在38次传代过程中，感染96-120小时后活力超过90%，在此期间活力提高，收获时间变得更加一致。在HeLa细胞中连续培养的刚地弓形虫可定期提供有活力的速殖子。证明HeLa来源的速殖子可用于染色试验，证实了该体外系统在综合医院实验室应用的潜力。
