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

  • 1Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea
  • 2The Center for Epidemic Preparedness, Korea Advanced Institute of Science and Technology (KAIST) Institute, Daejeon 34141, Korea


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.


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