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应用国产程序降温仪、[~3H]-TdR参入试验等研究了多种因素对小鼠脾淋巴细胞深低温保存的影响,并在此基础上建立起冻存该类细胞的最佳程序。结果表明,高浓度二甲亚砜对淋巴细胞毒性很大,但浓度在10％时对细胞的活力和功能无明显影响,且能有效地保护细胞免遭低温损伤;1℃/min降温保存效果最好,稍快速率降温时盛于冷冻塑料小管中的淋巴细胞功能恢复不及盛于HF冷冻袋中的细胞;复温后快速稀释或在冰浴中稀释都不利于淋巴细胞功能恢复;经Tris-NH_4Cl处理过的小鼠脾淋巴细胞对低温损伤的敏感性有些品系差异。用最佳程序冻存的脾淋巴细胞,在体内外测试中都显示出与冻存前相似的活力和功能,-196℃下贮存3个月也保存完好。
The effects of various factors on the cryopreservation of mouse splenic lymphocytes were studied using a domestic program thermometer, [~ 3H] -TdR incorporation test, and the best procedure for cryopreservation of such cells was established. The results showed that dimethyl sulfoxide (DMSO) was highly toxic to lymphocytes, but had no effect on the viability and function of cells at 10% concentration, and could effectively protect cells from hypothermia. The effect of cooling at 1 ℃ / min The best, slightly faster rate of cooling in plastic tubes filled with cryopreservation of lymphocytes function less than in the HF freezer cells; rapid rewarming after dilution or dilution in an ice bath are not conducive to lymphocyte function recovery; Tris There is some difference in the susceptibility of cold-damaged mice to splenic lymphocytes treated with -NH_4Cl. Spleen lymphocytes, which have been cryopreserved by the best procedure, showed similar viability and function to those before cryopreservation in both in vitro and in vivo tests and were also well preserved at -366C for 3 months.