AIDS Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) - Hospital Clinic, University of Barcelona, Barcelona, Spain.
Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.
J Immunol Methods. 2021 Nov;498:113133. doi: 10.1016/j.jim.2021.113133. Epub 2021 Sep 2.
The study of the effect of cryopreservation on the functionality of monocyte-derived dendritic cells (MDDCs) and dendritic cells (DCs) is essential for their use in different clinical applications such as DCs-based vaccines. Its full maturation and its optimal functionality are crucial for DCs based immunotherapy. In this study, we compared MDDCs derived from fresh and cryopreserved PBMCs in the aspects of phenotype and its effect on T cells at the level of proliferation and cytokine secretion. We pulsed MDDCs obtained from fresh and cryopreserved PBMCs with two different stimuli, CEF and SEA, and the expression maturation markers and cytokine secretion were analyzed. Our results showed that the cryopreservation had no effects in the phenotype of the MDDCs obtained, cell viability, maturation markers expression and/or cytokines secretion, independently whether MDDCs had been generated from fresh or cryopreserved PBMCs. Thus, this study suggests that the use of cryopreserved cells is a good method to keep the cells before use in immunotherapy, avoiding the variability within same individual due to severe blood draws. Even so, the interpretation and comparison of different results should be done considering the different cryopreservation techniques and assays, and their effects on PBMCs, specifically on MDDC and DC cells.
研究冷冻保存对单核细胞来源的树突状细胞（MDDC）和树突状细胞（DC）功能的影响对于它们在不同的临床应用中使用至关重要，例如基于 DC 的疫苗。其完全成熟及其最佳功能对于基于 DC 的免疫疗法至关重要。在这项研究中，我们比较了新鲜和冷冻保存的 PBMC 来源的 MDDC 在增殖和细胞因子分泌水平上对 T 细胞的表型及其影响。我们用两种不同的刺激物 CEF 和 SEA 脉冲处理从新鲜和冷冻保存的 PBMC 中获得的 MDDC，并分析了成熟标志物的表达和细胞因子的分泌。我们的结果表明，冷冻保存对获得的 MDDC 的表型、细胞活力、成熟标志物表达和/或细胞因子分泌没有影响，无论是从新鲜还是冷冻保存的 PBMC 中生成的 MDDC 都是如此。因此，这项研究表明，在免疫疗法之前使用冷冻保存的细胞是一种很好的方法，可以避免由于严重的血液采集而导致个体内的可变性。即便如此，对于不同结果的解释和比较应该考虑到不同的冷冻保存技术和检测方法，以及它们对 PBMC 特别是 MDDC 和 DC 细胞的影响。
