National-Local Associated Engineering Laboratory (Shenzhen) for Personalized Cellular Therapy, Nanshan, Shenzhen, Guangdong, China.
Shenzhen Beike Biotechnology Co., Ltd., Nanshan, Shenzhen, Guangdong, China.
Cryobiology. 2019 Feb;86:25-32. doi: 10.1016/j.cryobiol.2019.01.004. Epub 2019 Jan 7.
In this study, we compared three commercially available and two widely used CPAs for their ability of cryopreserving PBMCs. Similar survival (81.0%) and recovery rate (73.7%) were observed among cells using these five CPAs. However, all the cryopreserved PBMCs exhibited a significantly lower survival rate when compared with the fresh samples (94.3%). We further evaluated effector cell subpopulation and tumoricidal activity of PBMC-derived cytokine-induced killing (CIK) cells and natural killing (NK) cells. Similar and high survival (CIK: 88.6%; NK: 87.5%) and recovery (CIK: 99.5%; NK: 99.7%) rates were detected in CIK and NK cells prepared from cryopreserved PBMCs using the five CPAs. The CD3CD56 effector percentage (27.3%) of cryopreserved PBMC-derived CIK cells using the five different CPAs and their tumoricidal activities on melanoma CHL-1 cells (45.7%) and bladder cancer cell line T-24 (44.7%) were similar but significantly lower than those of the fresh PBMC-derived controls (effector: 30.7%; CHL-1: 84.2%; T-24: 82.2%). Cryopreserved PBMC-derived NK cells also exhibited similar tumoricidal activities (CHL-1: 73.8%; T-24: 71.9%) but was significantly lower than that of the fresh control group. We were not able to identify a specific CPA that performed superior than others in PBMC cryopreservation.
在这项研究中，我们比较了三种市售的和两种广泛使用的细胞冻存液在冻存 PBMCs 方面的能力。使用这五种细胞冻存液，细胞的存活率（81.0%）和回收率（73.7%）相似。然而，与新鲜样本（94.3%）相比，所有冻存的 PBMCs 的存活率都显著降低。我们进一步评估了 PBMC 来源的细胞因子诱导的杀伤（CIK）细胞和自然杀伤（NK）细胞的效应细胞亚群和细胞毒性活性。使用五种细胞冻存液，从冻存的 PBMC 中制备的 CIK 和 NK 细胞的存活率（CIK：88.6%；NK：87.5%）和回收率（CIK：99.5%；NK：99.7%）相似且较高。从冻存的 PBMC 中制备的 CIK 细胞中，使用五种不同的细胞冻存液的 CD3CD56 效应细胞比例（27.3%）及其对黑色素瘤 CHL-1 细胞（45.7%）和膀胱癌 T-24 细胞系（44.7%）的细胞毒性活性相似，但显著低于新鲜 PBMC 衍生对照物的活性（效应细胞：30.7%；CHL-1：84.2%；T-24：82.2%）。冻存的 PBMC 衍生的 NK 细胞也表现出相似的细胞毒性活性（CHL-1：73.8%；T-24：71.9%），但显著低于新鲜对照组。我们未能确定一种在 PBMC 冻存方面表现优于其他的特定细胞冻存液。
