Single-cell transcriptome conservation in a comparative analysis of fresh and cryopreserved human skin tissue: pilot in localized scleroderma

Emily Mirizio, Tracy Tabib, Xiao Wang, Wei Chen, Christopher Liu, Robert Lafyatis, Heidi Jacobe, Kathryn S. Torok

Research output: Contribution to journalArticlepeer-review

Abstract

Background: The purpose of this study was to assess variability in cell composition and cell-specific gene expression in the skin of patients with localized scleroderma (LS) utilizing CryoStor® CS10 in comparison to RPMI to produce adequate preservation of tissue samples and cell types of interest for use in large-scale multi-institutional collaborations studying localized scleroderma and other skin disorders. Methods: We performed single-cell RNA sequencing on paired skin biopsy specimens from 3 patients with LS. Each patient with one sample cryopreserved in CryoStor® CS10 and one fresh in RPMI media using 10× Genomics sequencing. Results: Levels of cell viability and yield were comparable between CryoStor® CS10 (frozen) and RPMI (fresh) preserved cells. Furthermore, gene expression between preservation methods was collectively significantly correlated and conserved across all 18 identified cell cluster populations. Conclusion: Comparable cell population and transcript expression yields between CryoStor® CS10 and RPMI preserved cells support the utilization of cryopreserved skin tissue in single-cell analysis. This suggests that employing standardized cryopreservation protocols for the skin tissue will help facilitate multi-site collaborations looking to identify mechanisms of disease in disorders characterized by cutaneous pathology.

Original languageEnglish (US)
Article number263
JournalArthritis Research and Therapy
Volume22
Issue number1
DOIs
StatePublished - Dec 2020

Keywords

  • Cryopreservation
  • Localized scleroderma
  • Morphea
  • Pediatric rheumatology
  • Single-cell RNA sequencing
  • Transcriptome expression

ASJC Scopus subject areas

  • Rheumatology
  • Immunology and Allergy
  • Immunology

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