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KMID : 0923620200200040034
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Immune Network
2020 Volume.20 No. 4 p.34 ~ p.34
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Effects of Cryopreservation and Thawing on Single-Cell Transcriptomes of Human T Cells
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Lee Jeong-Seok
Yi Ki-Jong
Ju Young-Seok
Shin Eui-Cheol
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Abstract
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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.
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KEYWORD
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Cryopreservation, Single-cell analysis, Transcriptome, T-lymphocytes, regulatory
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