CRYOPRESERVATION OF SCHISTOSOMULES OF SCHISTOSOMA-MANSONI IN QUANTITY
CRYOPRESERVATION OF SCHISTOSOMULES OF SCHISTOSOMA-MANSONI IN QUANTITY
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DOI：
10.4269/ajtmh.1984.33.116
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发表时间：
1984-01-01
影响因子：
3.3
通讯作者：
LEEF, JL
中科院分区：
文献类型：
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作者：
STIREWALT, M;LEWIS, FA;LEEF, JL
大量的S。将曼氏疟原虫孢子在35%乙醇中冷冻保存，并尝试在解冻后改善其形态和感染性。几个变量影响解冻生物体的外观。其中包括：用于将尾蚴转化为囊泡的特定体外方法;向解冻培养基中添加5-羟色胺;改变解冻温度;以及在评估囊泡状况之前培养囊泡延长的解冻后时间。 改变转化前尾蚴的羽化后年龄或冷冻悬浮液中生物体的浓度对其存活没有影响。虽然许多解冻的溶酶体的损害变得明显，只有在延长体外培养，加入胎牛血清解冻培养基通常延缓这种恶化。当注射到小鼠体内时，5%的冷冻保存的精子成熟为蠕虫，占未冷冻精子成熟值的71%。这些研究确定了在液氮中成功保存大量的微囊体（1 ml体积中可达500，000个）的重要条件。
Large quantities of S. mansoni schistosomules were cryopreserved in 35% ethanediol, and attempts were made to improve their morphology and infectivity after thawing. Several variables affected the appearance of the thawed organisms. These included: the particular in vitro method used to transform cercariae to schistosomules; addition of serotonin to the thawing medium; changing the thawing temperature; and culturing schistosomules for extended post thaw times before assessing their condition. Varying either the postemergence age of the cercariae before transformation or the concentration of organisms in the freezing suspension had no effect on their survival. Although damage to many of the thawed schistosomules became apparent only upon extended in vitro cultivation, the addition of fetal calf serum to the thawing medium usually retarded this deterioration. When injected into mice, .apprx. 5% of cryopreserved schistosomules matured to adult worms, representing 71% of the value for maturation of unfrozen schistosomules. These studies define conditions of importance for the successful preservation of very large quantities of schistosomules (up to 500,000 in 1 ml volume) in liquid N2.