Immune Netw.  2020 Aug;20(4):e34. 10.4110/in.2020.20.e34.

Effects of Cryopreservation and Thawing on Single-Cell Transcriptomes of Human T Cells

• Lee JS
• Yi K
• Ju YS
• Shin EC

• KMID: 2505678
• DOI: http://doi.org/10.4110/in.2020.20.e34

Abstract

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.

Keyword

Cryopreservation; Single-cell analysis; Transcriptome; T-lymphocytes, regulatory