Weiner R S
J Immunol Methods. 1976;10(1):49-60. doi: 10.1016/0022-1759(76)90006-5.
Human lymphocytes can be cryopreserved for the purpose of performing in vitro tests of cellular immunity. A systematic study of the conditions for freezing and recovering cells has shown that there is wide flexibility in cell concentrations of cryoprotective agent and the temperature and rate of dilution however, have definable optima. Cells frozen and thawed under optimal conditions retain their reactivity in MLC and can be used for sequential studies of immune responsiveness. Approximately 70% of viable and functional cells are recovered when the cells are frozen in 7.5--12.5% DMSO and no further cryoprotection is discernable when up to 50% serum is added to the freezing media. The temperature and rate of dilution are critical only in that cells diluted rapidly in THE COLD (10-fold in 2 min at 0 degrees C) are less responsive in MLC than are cells diluted slowly (10-fold in 10 min) in the cold or slowly or rapidly at room temperature.
为了进行细胞免疫的体外测试，可以对人类淋巴细胞进行冷冻保存。一项关于细胞冷冻和复苏条件的系统研究表明，冷冻保护剂的细胞浓度具有很大的灵活性，然而，稀释温度和速率具有明确的最佳值。在最佳条件下冷冻和解冻的细胞在混合淋巴细胞培养（MLC）中保留其反应性，并且可用于免疫反应性的连续研究。当细胞在7.5%-12.5%的二甲基亚砜（DMSO）中冷冻时，约70%的存活且有功能的细胞得以复苏，并且当向冷冻培养基中添加高达50%的血清时，未发现进一步的冷冻保护作用。稀释温度和速率至关重要，仅在于在低温下快速稀释的细胞（在0℃下2分钟内稀释10倍）在MLC中的反应性低于在低温下缓慢稀释（10分钟内稀释10倍）或在室温下缓慢或快速稀释的细胞。
