Project description:The pig skin architecture and physiology are similar to these of humans. Thus, the pig model is valuable for studying skin biology and testing therapeutics for skin diseases. The single-cell RNA sequencing technology allows quantitatively analyzing cell types, cell 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. To maximize the use of pig skin as a model system to study skin biology, we described a robust method for isolating and cryo-preserving pig 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. In addition, we showed that the subsequent single cells could be cryopreserved using 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 will be very valuable for the skin research scientific community.
Project description:To study the development of pig facial skin after birth, we use the facial skin tissues of healthy Chenghua sows as experimental materials. we then performed gene expression profiling analysis using data obtained from RNA-seq of pig facial skin tissues at four time points.
Project description:Skin is the largest barrier organ with complex structure and function. We investigated the transcriptional dynamics of pig skin during development and aging at single-cell resolution. Using single-cell RNA sequencing, we profiled the transcriptomes of 443,529 skin cells derived from 30 individuals of 10 developmental time points staged between 56 days of gestation and 7 years old. Our analysis identified the dynamic changes in the cellular transcriptional profile of eight skin cell types at different developmental stages, exhibiting time-dependent skin cell heterogeneity.
Project description:Study to optimize our protocol for isolating RNA from skin biopsies from (hairless) SKH mice using different-diameter biopsy punches. Some mice were also treated with UVB radiation to check its effect on RNA yield.