Campbell S J, Ingram P R, Roberts C W, Henriquez F L
School of Engineering and Science, University of the West of Scotland, Paisley PA1 2BE, Scotland.
Parasitology. 2008 Oct;135(12):1401-5. doi: 10.1017/S0031182008005003.
Several conditions that allow the preservation, storage and rapid, efficient recovery of viable Acanthamoeba castellanii organisms were investigated. The viability of trophozoites (as determined by time to confluence) significantly declined over a period of 12 months when stored at -70 degrees C using dimethyl sulfoxide (DMSO; 5 or 10%) as cryopreservant. As A. castellanii are naturally capable of encystment, studies were undertaken to determine whether induced encystment might improve the viability of organisms under a number of storage conditions. A. castellanii cysts stored in the presence of Mg2+ at 4 degrees C remained viable over the study period, although time to confluence was increased from approximately 8 days to approximately 24 days over the 12-month period. Storage of cysts at -70 degrees C with DMSO (5 or 10%) or 40% glycerol, but not 80% glycerol as cryopreservants increased their viability over the 12-month study period compared with those stored at room temperature. Continued presence of Mg2+ in medium during storage had no adverse effects and generally improved recovery of viable organisms. The present study demonstrates that A. castellanii can be stored as a non-multiplicative form inexpensively, without a need for cryopreservation, for at least 12 months, but viability is increased by storage at -70 degrees C.
研究了几种能够保存、储存并快速高效复苏存活的卡氏棘阿米巴生物体的条件。当使用二甲基亚砜（DMSO；5%或10%）作为冷冻保护剂在-70℃储存时，滋养体的活力（通过汇合时间确定）在12个月内显著下降。由于卡氏棘阿米巴天然具有形成包囊的能力，因此开展了研究以确定诱导形成包囊是否可能在多种储存条件下提高生物体的活力。在4℃含有Mg2+的情况下储存的卡氏棘阿米巴包囊在研究期间保持存活，尽管在12个月期间汇合时间从大约8天增加到了大约24天。与在室温下储存的包囊相比，在-70℃使用DMSO（5%或10%）或40%甘油而非80%甘油作为冷冻保护剂储存包囊，在12个月的研究期间提高了它们的活力。储存期间培养基中持续存在Mg2+没有不利影响，并且总体上提高了存活生物体的复苏率。本研究表明，卡氏棘阿米巴可以作为非增殖形式廉价储存，无需冷冻保存，至少可保存12个月，但在-70℃储存可提高活力。
