Sears H F, Rosenberg S A
J Natl Cancer Inst. 1977 Feb;58(2):183-7. doi: 10.1093/jnci/58.2.183.
Human lymphocytes were harvested from normal volunteer donors and cryopreserved. Various concentrations of dimethyl sulfoxide and human serum in the cryoprotective media were evaluated to optimize the recovery and function of viable cells. Multiple samples were then drawn from volunteers over a number of days, processed individually, and used in mitogen stimulation assays. These cells were also cryopreserved, immediately thawed, and cultured simultaneously with the fresh cells. In all instances fresh and cryopreserved lymphocytes exhibited similar levels of stimulation by mitogens. Cryopreserved cells from these sequential bleedings were then recovered and cultured simultaneously in mitogen stimulation assays. The results obtained in these assays with cryopreserved cell had less day-to-day variagion than those with cells cultured individually. Coefficients of variance of individual cultures of mitogen stimulation assays were reduced from 26-59% to 5-17% by use of cryopreserved cells. The findings suggested that use of cryopreservation techniques decreases the variability of cellular immune assays and thus alows more accurate longitudinal study of the immune competence of patients.
从正常志愿者供体采集人淋巴细胞并进行冷冻保存。评估冷冻保护培养基中不同浓度的二甲基亚砜和人血清，以优化活细胞的回收率和功能。然后在数天内从志愿者身上采集多个样本，分别进行处理，并用于有丝分裂原刺激试验。这些细胞也进行冷冻保存，立即解冻，并与新鲜细胞同时培养。在所有情况下，新鲜的和冷冻保存的淋巴细胞对有丝分裂原的刺激水平相似。然后回收这些连续采血得到的冷冻保存细胞，并在有丝分裂原刺激试验中同时进行培养。在这些试验中，使用冷冻保存细胞得到的结果比单独培养细胞的结果具有更小的每日变化。通过使用冷冻保存细胞，有丝分裂原刺激试验的个体培养物的变异系数从26 - 59%降至5 - 17%。这些发现表明，使用冷冻保存技术可降低细胞免疫测定的变异性，从而允许对患者的免疫能力进行更准确的纵向研究。
