Mavin S, Evans R, Chatterton J M W, Ashburn D, Joss A W L, Ho-Yen D O
Scottish Toxoplasma Reference Laboratory, Raigmore Hospital, Inverness IV2 3UJ, UK.
Br J Biomed Sci. 2003;60(4):217-20. doi: 10.1080/09674845.2003.11783702.
This study aims to increase the efficiency of continuous growth of Toxoplasma gondii in HeLa cells from tachyzoite stocks frozen in liquid nitrogen. Freezing and retrieval of tachyzoites for continuous cell culture requires more stringent protocols than those published for animal culture. The freezing and retrieval conditions are optimised so that a quality harvest (> or = 1 x 10(6) tachyzoites/mL, > or = 90% viability) can be produced using T. gondii recovered from liquid nitrogen as fast and reliably as possible. Retrieval success rate increased from 36% to 100%. An improved freezing procedure using chilled reagents and freshly harvested parasites, and adoption of an effective recovery protocol with retrieval of 3 x 10(7) tachyzoites into 75 cm2 flasks, change of maintenance media after six hours and subsequent blind passage all contributed to this success. The result is faster and more dependable production of T. gondii for diagnostic and experimental use.
本研究旨在提高从液氮中冷冻保存的速殖子库存中连续培养刚地弓形虫的效率。用于连续细胞培养的速殖子冷冻和复苏需要比已发表的动物培养方案更为严格的规程。对冷冻和复苏条件进行了优化，以便能尽可能快速且可靠地利用从液氮中复苏的刚地弓形虫获得优质收获物（≥1×10⁶个速殖子/毫升，≥90%的活力）。复苏成功率从36%提高到了100%。使用预冷试剂和新鲜收获的虫体改进冷冻程序，采用将3×10⁷个速殖子接种到75平方厘米培养瓶中、6小时后更换维持培养基并随后进行盲传的有效复苏方案，这些都促成了这一成功。结果是能更快且更可靠地生产用于诊断和实验的刚地弓形虫。
