Shenzhen Beike Biotechnology Co., Ltd., Nanshan, Shenzhen, Guangdong, China.
National-Local Associated Engineering Laboratory for Personalized Cellular Therapy, Nanshan, Shenzhen, Guangdong, China.
Cell Tissue Bank. 2020 Dec;21(4):631-641. doi: 10.1007/s10561-020-09857-w. Epub 2020 Aug 18.
Peripheral blood mononuclear cells are widely used as source material for anticancer immunotherapies. The conventional cryopreservation method for peripheral blood mononuclear cells is time-consuming and expansive, which involves controlled rate freezing followed by storage in liquid nitrogen. Instead, the convenient uncontrolled rate freezing cryopreservation method had been reported successfully in peripheral blood hematopoietic stem cells and peripheral blood progenitor cells. Therefore, we hypothesized that uncontrolled rate freezing cooling method maybe also applied to peripheral blood mononuclear cells cryopreservation. In this study, we evaluated the performance of uncontrolled rate freezing and controlled rate freezing cooling methods through cell recovery rate, viability, differentiation potential into cytokine-induced killer cells and the cellular properties of the cultured cytokine-induced killer cells. The results showed similar post-thaw viability and recovery rate in both controlled rate freezing and uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells. Importantly, the uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells exhibited higher growth ratio and earlier cell clustering during ex-vivo cytokine-induced killer cell culture than the controlled rate freezing ones. These two groups of expanded cytokine-induced killer cells also exhibited similar effector cell subset ratio and tumoricidal activity. In general, the performance of cryopreserved peripheral blood mononuclear cells using uncontrolled rate freezing cooling method, with the commercial cryoprotective agent CellBanker 2, was equal or better than the controlled rate freezing method. Our study implied that the combined use of cryoprotective agent CellBanker 2 and uncontrolled rate freezing could be a convenient cryopreservation method for peripheral blood mononuclear cells.
外周血单个核细胞广泛用作抗癌免疫疗法的来源材料。外周血单个核细胞的传统冷冻保存方法既耗时又昂贵，涉及控制速率冷冻，然后储存在液氮中。相反，已经成功报道了外周血造血干细胞和外周血祖细胞的方便的非控制性速率冷冻保存方法。因此，我们假设非控制性速率冷冻冷却方法也可应用于外周血单个核细胞的冷冻保存。在这项研究中，我们通过细胞回收率、活力、分化为细胞因子诱导的杀伤细胞的潜能以及培养的细胞因子诱导的杀伤细胞的细胞特性来评估非控制性速率冷冻和控制性速率冷冻冷却方法的性能。结果表明，在控制性速率冷冻和非控制性速率冷冻保存的外周血单个核细胞中，解冻后的活力和回收率相似。重要的是，与控制性速率冷冻保存的外周血单个核细胞相比，非控制性速率冷冻保存的外周血单个核细胞在体外细胞因子诱导的杀伤细胞培养中表现出更高的生长比和更早的细胞聚集。这两组扩增的细胞因子诱导的杀伤细胞也表现出相似的效应细胞亚群比例和杀肿瘤活性。总的来说，使用非控制性速率冷冻冷却方法（使用商业细胞保护剂 CellBanker 2）冷冻保存外周血单个核细胞的性能与控制性速率冷冻方法相当或更好。我们的研究表明，细胞保护剂 CellBanker 2 和非控制性速率冷冻的联合使用可能是外周血单个核细胞的一种方便的冷冻保存方法。
