Chatterton Jean M W, Evans R, Ashburn D, Joss A W L, Ho-Yen D O
Scottish Toxoplasma Reference Laboratory, Microbiology Department, Raigmore Hospital, Highland Acute Hospitals NHS Trust, Old Perth Road, Iverness IV2 3UJ, Scotland, UK.
J Microbiol Methods. 2002 Nov;51(3):331-5. doi: 10.1016/s0167-7012(02)00101-x.
The aim of this study was to develop a culture method for Toxoplasma gondii that could provide fresh viable tachyzoites as and when required. When T. gondii was continuously maintained in HeLa cell cultures at 37 degrees C, the time to harvest varied from 48 to 144 h. Tachyzoite yields of > or = 1 x 10(6)/ml and > or = 90% viable were obtained from 519/882 (58.8%) cultures and 120/155 (77.4%) harvests were successfully used in the dye test. When cultures were transferred from 37 to 25 degrees C when maximally infected (48-54-h post-infection), they could be stabilised and tachyzoites could be harvested as required, up to 168 h later. When harvested from 25 degrees C, significantly more cultures 783/811 (96.5%) produced tachyzoite yields > or = 1 x 10(6)/ml > or = 90% viable (p < 0.001). Tachyzoite quality also significantly improved and 206/224 harvests (91.9%) (p < 0.001) were successfully used in the dye test. We have demonstrated that tachyzoites can be maintained at dye test quality for at least 7 days in HeLa cultures at 25 degrees C. The system is flexible and robust and provides a means whereby tachyzoites of standard quality can be stored for use in experimental models as and when required.
本研究的目的是开发一种用于刚地弓形虫的培养方法，该方法能够在需要时提供新鲜的活速殖子。当刚地弓形虫在HeLa细胞培养物中于37℃连续传代培养时，收获所需的时间为48至144小时。从519/882（58.8%）的培养物中获得了产量≥1×10⁶/ml且活力≥90%的速殖子，120/155（77.4%）的收获物成功用于染色试验。当培养物在感染达到最大程度时（感染后48 - 54小时）从37℃转移至25℃，它们可以被稳定下来，并且可以根据需要收获速殖子，最长可在168小时后收获。当从25℃收获时，显著更多的培养物783/811（96.5%）产生了产量≥1×10⁶/ml且活力≥90%的速殖子（p < 0.001）。速殖子的质量也显著提高，206/224次收获（91.9%）（p < 0.001）成功用于染色试验。我们已经证明，速殖子在25℃的HeLa培养物中可以保持染色试验所需质量至少7天。该系统灵活且稳定，提供了一种方法，通过该方法可以将标准质量的速殖子储存起来，以便在需要时用于实验模型。
