Weed R G, Jenkins E C
Exp Cell Biol. 1979;47(5):351-9. doi: 10.1159/000162954.
It was previously shown by the authors that human whole blood was cryoprotected by dimethyl sulfoxide (DMSO) for use as inoculum in short-term lymphocyte cultures. That procedure utilized a fast postthaw dilution where the cryoprotective agent was diluted in two steps. In this paper it is shown that a five-step 'slow' dilution technique is much superior to the two-step 'fast' dilution procedure, which significantly revises our original report of 1972. This method has strikingly improved the degree of blastogenic response to phytohemagglutinin as measured by 3H-thymidine incorporation. The increase in amount of tritium incorporation was seen in cultures which were protected with either DMSO or glycerol. A similar effect was observed in unfrozen control cultures exposed to either cryoprotectant.
作者之前曾表明，人全血可用二甲基亚砜（DMSO）进行冷冻保护，以用作短期淋巴细胞培养中的接种物。该程序采用快速解冻后稀释法，其中冷冻保护剂分两步稀释。本文表明，五步“缓慢”稀释技术远优于两步“快速”稀释程序，这显著修正了我们1972年的原始报告。通过3H-胸腺嘧啶核苷掺入量测定，该方法显著提高了对植物血凝素的增殖反应程度。在用DMSO或甘油保护的培养物中，氚掺入量均有所增加。在暴露于任何一种冷冻保护剂的未冷冻对照培养物中也观察到了类似的效果。
