Isolating and cryopreserving pig skin cells for single-cell RNA sequencing study
Abstract
The pig skin architecture and physiology are similar to those of humans. Thus, the pig model is very valuable for studying skin biology and testing therapeutics. The single-cell RNA sequencing (scRNA-seq) technology allows quantitatively analyzing cell types, compositions, states, signaling, and receptor-ligand interactome at single-cell resolution and at high throughput. scRNA-seq has been used to study mouse and human skins. However, studying pig skin with scRNA-seq is still rare. A critical step for successful scRNA-seq is to obtain high-quality single cells from the pig skin tissue. Here we report a robust method for isolating and cryopreserving pig skin single cells for scRNA-seq. We showed that pig skin could be efficiently dissociated into single cells with high cell viability using the Miltenyi Human Whole Skin Dissociation kit and the Miltenyi gentleMACS Dissociator. Furthermore, the obtained single cells could be cryopreserved using 90% FBS + 10% DMSO without causing additional cell death, cell aggregation, or changes in gene expression profiles. Using the developed protocol, we were able to identify all the major skin cell types. The protocol and results from this study are valuable for the skin research scientific community.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Feb 17, 2022
- Source ID
- 10.1371/journal.pone.0263869
Entities
People
- Carlos P. Jara
- Chi Zhang
- Eliana P. Araújo
- Li Han
- Marc Libault
- Mark A. Carlson
- Ou Wang
- Rafał K. Wóycicki
- Sandra Thibivilliers
- William H. Velander
- Xinran Wu
- Yu Shi
- Yuguo Lei
Organizations
- Coordenação de Aperfeicoamento de Pessoal de Nível Superior
- Foundation for the National Institutes of Health
- Nebraska Department of Health and Human Services
- Pennsylvania State University
- United States Army
- United States Department of Defense
- University of Nebraska system
- University of Nebraska–Lincoln