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 language | English (US) |
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Article number | 263 |
Journal | Arthritis Research and Therapy |
Volume | 22 |
Issue number | 1 |
DOIs | |
State | Published - Dec 2020 |
Keywords
- Cryopreservation
- Localized scleroderma
- Morphea
- Pediatric rheumatology
- Single-cell RNA sequencing
- Transcriptome expression
ASJC Scopus subject areas
- Rheumatology
- Immunology and Allergy
- Immunology