Ham P J, James E R, Bianco A E
Trans R Soc Trop Med Hyg. 1982;76(6):758-67. doi: 10.1016/0035-9203(82)90101-8.
A technique has been developed for separating viable and non-viable Onchocerca lienalis microfilariae (mff) by passing a suspension of the parasites through a column of DEAE cellulose. A proportion (between 42% and 87%) of normally motile parasites, whether unfrozen or cryopreserved, passed through the cellulose columns and retained their viability. Non-motile and sluggish mff were retained by the cellulose. The proportion of viable cryopreserved mff was greater after passage through the column as assessed by motility, migration in mice and development in the insect vector. Numbers of cryopreserved mff eluted through the columns peak quickly at a relatively high level after about 20 ml of eluate had been passed through, whereas normal unfrozen parasites rose and fell in numbers more slowly. By using sodium chloride gradients and buffers of differing conductivity it was concluded that the separation of the mff is probably due to net charge changes as well as motility. It is believed that this technique offers considerable promise as a tool to provide populations of mff of more uniform viability following freezing and thawing with existing cryopreservation techniques.
已开发出一种技术，通过使寄生虫悬浮液通过DEAE纤维素柱来分离存活和不存活的链尾盘尾丝虫微丝蚴（mff）。无论未冷冻还是冷冻保存，一定比例（42%至87%）的正常活动的寄生虫通过纤维素柱并保持其活力。不活动和行动迟缓的mff被纤维素保留。通过活动能力、在小鼠体内的迁移和在昆虫媒介中的发育评估，冷冻保存的mff通过柱后的存活比例更高。约20毫升洗脱液通过后，通过柱洗脱的冷冻保存mff数量在相对较高水平迅速达到峰值，而正常未冷冻的寄生虫数量上升和下降则较为缓慢。通过使用氯化钠梯度和不同电导率的缓冲液得出结论，mff的分离可能是由于净电荷变化以及活动能力。据信，作为一种工具，该技术有望在用现有冷冻保存技术进行冷冻和解冻后，提供活力更均匀的mff群体。
