Project description:Cells were isolated from the back skin of 1-day old chicks after plucking down feathers. Cells were dissociated and enriched for epidermal cells according to a published protocol (PMID: 33157095). Single‐cell separation (10X Genomics Chromium), library preparation (10X Genomics Cell Ranger, version 3.0.2), sequencing (Illumina HiSeq 3000/4000) and data analysis (Seurat Version 3, Satija Lab, NY) were performed according to a published protocol (PMID: 31930613).
Project description:We sequenced single cells with the 10x platform of chicken spleens from chickens genetically resistant and genetically susceptible to Marek's disease, with both infected individuals and uninfected controls from both lines
Project description:Single-cell RNA sequencing is transforming how we understand skin immunology, but previous human skin single-cell RNA sequencing data included only a small fraction of inflammatory cells among the overall cell population, such that functional subsets may be difficult to ascertain. We have overcome these obstacles by harvesting inflammatory cells emigrating from a half of 6 mm punch biopsy skin after 48-hour incubation in culture medium without any enzyme, and then analyzing the harvested cells with single-cell RNA sequencing. By this strategy, we obtained single-cell RNA sequencing data of 24,354 cells (leukocytes 46.0%, keratinocytes 49.6%, and melanocytes 2.4%) from 13 human psoriasis skin and 5 healthy volunteer skin. Unsupervised clustering identified NK cells, CD161+ T-cells, CD8+ T-cells, CD4+ T-cells, regulatory T-cells, mature & semimature dendritic cells, melanocytes, and keratinocytes in different layers - S. (Stratum) corneum, S. granulosum, S. spinosum, and S. basale. To understand psoriasis immunopathogenesis at single-cell levels, we compared gene expression between psoriasis cells vs. control cells within each inflammatory cell subtype clusters.
Project description:Despite the recent application of single-cell RNA-sequencing to aspects of mouse skin biology, the full cellular heterogeneity of the mouse skin (including both epidermis and skin stroma) and its relationship with the hair cycle is still uncharted. In order to systematically compare the cellular composition of mouse skin during rest and hair growth, we created single-cell RNA-sequencing libraries from full thickness mouse skin cell suspensions sampled during anagen (5w) and telogen (9w).
Project description:Ma-Huang chicken as a high-quality broiler is one of the most popular chickens in the frozen chicken market. However, some chicken may instead have white or lighter skin, which directly causes economic losses every year. To obtain better insight into the molecular mechanisms associated with the process of pigmentation of yellow-skinned broilers reared under intensive conditions, a total of six-hundred Ma-Huang chickens was randomly selected in a single slaughterhouse, the color measurements were carried out on both cloaca(alive) and five different part after slaughtering adopting the L* a* b* system and using a 3nh-NH310 colorimeter, color values from areas of the chicken skin pear each image automatically retrieved by MATLAB, production and slaughtering traits were also measured, comparative transcriptomic analysis of high yellowness(s_deep) versus low yellowness(s_light) skin was performed using the Illumina Hiseq 4000. Average values of the cloaca(alive), cloaca (hair removal), thigh, shank and abdominal fat were 8.98, 7.66, 2.62, 7.29 and 12.86, respectively. The better production and slaughtering traits were observed in higher skin yellowness chicken. Yellowness values of the cloaca(alive) and after slaughtering were significantly correlated (p < 0.05), suggesting that the color of after slaughtering evaluation may be carried out on cloaca(alive). A total of 19061 unigenes were assembled from the reads obtained from the skin of two groups, 882 unigenes were differentially expressed between s_deep and s_light (Fold change ≥ 2, Adjusted P <= 0.001), 612 that up-regulated and 270 that down-regulated genes in s_deep skin, as compared with s_light skin. Twelve promising candidate genes may play an important role in the pigmentation of chicken skin, i.e. GPR143, PMEL, TYR, CYP11A1, TECRL, ACACB, TLR2B, ALDH1A3, FHL2, TECRL, DUOX2, SMOC1 were included. Furthermore, some important functional pathways were revealed, such as the biological process, cellular component and molecular function, which appear to be much activity in skin pigmentation. Our data provide a valuable resource for identifying genes whose functions are critical to skin pigmentation, facilitate understanding the molecular mechanisms of the skin color variation on yellow-skinned broiler chickens under commercial conditions, accelerate the molecular selection of the specific strain on consistent skin colors which allow reduction in pigment use to achieve the skin acceptance by the consumer.