Cryopreservation and thawing of PBMCs are inevitable processes in expanding the scale of experiments in human immunology. Here, we carried out a fundamental study to investigate the detailed effects of PBMC cryopreservation and thawing on transcriptomes. We sorted Tregs from fresh and cryopreserved/thawed PBMCs from an identical donor and performed single-cell RNA-sequencing (scRNA-seq). We found that the cryopreservation and thawing process minimally affects the key molecular features of Tregs, including FOXP3. However, the cryopreserved and thawed sample had a specific cluster with up-regulation of genes for heat shock proteins. Caution may be warranted in interpreting the character of any cluster of cells with heat shock-related properties when cryopreserved and thawed samples are used for scRNA-seq.

中文翻译：

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冷冻保存和解冻对人 T 细胞单细胞转录组的影响

PBMC 的冷冻保存和解冻是扩大人类免疫学实验规模的必然过程。在这里，我们进行了一项基础研究，以调查 PBMC 冷冻保存和解冻对转录组的详细影响。我们从来自同一供体的新鲜和冷冻保存/解冻的 PBMC 中分选了 Treg，并进行了单细胞 RNA 测序 (scRNA-seq)。我们发现冷冻保存和解冻过程对 Tregs 的关键分子特征的影响最小，包括 FOXP3。然而，冷冻保存和解冻的样品有一个特定的簇，热休克蛋白的基因上调。当冷冻保存和解冻的样品用于 scRNA-seq 时，在解释具有热休克相关特性的任何细胞簇的特征时可能需要谨慎。