Department of Clinical Immunology, Blood Bank, Rigshospitalet, 2100, Copenhagen, Denmark.
Department of Clinical Immunology, Blood Bank, Rigshospitalet, 2100, Copenhagen, Denmark.
Transfus Apher Sci. 2021 Aug;60(4):103138. doi: 10.1016/j.transci.2021.103138. Epub 2021 Apr 16.
T cell based treatments in the setting of allogenic haematopoietic stem cell transplantation (HSCT) have been used for decades. In addition, the use of chimeric antigen receptor (CAR) T cells has been introduced as a promising cancer immunotherapy. A prerequisite for many of these treatments is the ability to cryopreserve the cells safely and efficiently. In the present study, we compared freezing media combinations containing pentaisomaltose and 1-2 % DMSO (PIM1 and PIM2, respectively) to 10 % DMSO and commercially available cryosolutions (CS2 and CS10, Cryostor® containing 2 and 10 % DMSO, respectively) for cryopreservation of T cells. T cells isolated from buffy coats from healthy donors were cryopreserved with different freezing media and analysed for 1) viability immediately post-thaw and the following 24 h, 2) recovery, 3) proliferative potential and 4) migration towards a gradient of SDF-1α. The results showed that PIM2 was superior to 10 % DMSO and comparable to CS10 when assessing viability. Furthermore, the results indicated that the T cells cryopreserved with 10 % DMSO showed the lowest proliferative potential. The expression levels of CXCR3, CXCR4 and VLA-4 were similar in T cells independent of the freezing media used; however, T cells cryopreserved with PIM2 demonstrated the highest migratory potential. In summary, the combination of pentaisomaltose and 1-2 % DMSO improves the cryoprotective properties compared to 10 % DMSO while achieving comparable results with CS10 and even showing improved migration towards SDF-1α. Thus, our results show promising potential for pentaisomaltose in combination with low amounts of DMSO for the cryopreservation of T cells.
T 细胞为基础的治疗在同种异体造血干细胞移植（HSCT）的背景下已经使用了几十年。此外，嵌合抗原受体（CAR）T 细胞的使用已经作为一种有前途的癌症免疫疗法被引入。这些治疗方法中的许多都需要能够安全有效地冷冻保存细胞。在本研究中，我们比较了含有五异麦芽糖和 1-2%二甲基亚砜（分别为 PIM1 和 PIM2）的冷冻液组合与 10%二甲基亚砜和市售冷冻液（CS2 和 CS10，分别含有 2%和 10%二甲基亚砜，Cryostor®），用于 T 细胞的冷冻保存。从健康供者的血涂片分离的 T 细胞用不同的冷冻液冷冻保存，并分析了 1）解冻后立即和接下来 24 小时的活力，2）回收率，3）增殖潜能和 4）向 SDF-1α梯度的迁移。结果表明，在评估活力方面，PIM2 优于 10%二甲基亚砜，与 CS10 相当。此外，结果表明，用 10%二甲基亚砜冷冻保存的 T 细胞表现出最低的增殖潜能。T 细胞冷冻保存所用的冷冻液对 CXCR3、CXCR4 和 VLA-4 的表达水平没有影响；然而，用 PIM2 冷冻保存的 T 细胞显示出最高的迁移潜能。总之，与 10%二甲基亚砜相比，五异麦芽糖和 1-2%二甲基亚砜的组合提高了冷冻保护性能，同时与 CS10 取得了相当的结果，甚至显示出对 SDF-1α的迁移能力有所提高。因此，我们的结果表明，五异麦芽糖与低剂量二甲基亚砜结合在 T 细胞冷冻保存方面具有很大的潜力。
