Project description:To obtain a large representation of non-immune skin-resident cells, we profiled CD45-negative cells from a digested skin sample taken from a human female
Project description:In this study, we analysed early embryonic skin development (mus musculus; C57BL/6J) at the transcriptional level. Major questions concerned the cell type composition of early embryonic skin, and the emergence of transcriptional heterogeneity among epithelial and stromal precursor cells. Cells were isolated from embryonic dorsal skin and randomly sequenced (scRNA-Seq using 10X Genomics v2) without any cell sorting. Data from three embryonic time points (E12.5, E13.5, and E14.5) was integrated and compared to obtain a better understanding of the dynamics of early skin development.
Project description:Epcam-positive cells from fetal intestinal epithelium were FACS sorted and processed using 10X genomics to characterise the differentiation of fetal intestinal cells at E16.5.
Project description:Cells were generated for a single cell atlas of adult human skin to dissect the cellular and molecular organisation underpinning human skin immune barrier function. Surplus skin from breast reconstruction surgery was collected (n=3), then the top 200 μm layer was taken. Epidermis was separated from the dermis, and both layers were separately digested. Single cells were FACS sorted into eight categories, then loaded onto a 10x Genomics Chromium Controller, and sequenced on an Illumina HiSeq 4000.
Project description:We performed an inter-species comparison of murine spermatogenesis using the inbred strains C57B6J, CAST/EiJ and CAROLI/EiJ to investigate the cell type-specific evolution of gene expression levels among closely related species. We also used F1 crosses of C57B6J and CAST/EiJ to investigate context-specific regulatory effects on gene expression in cis and trans by measuring allele-specific expression (Goncalves et al. 2012; Wittkopp et al. 2021). To this end, single-cell RNA-Sequencing data was generated from dissociated testicular tissue in each mouse strain using the 10x Genomics scRNA-Seq platform.
Project description:Single cell RNA-seq of SVZ lateral ventricle walls from non-irradiated mice brains and 5 days post-irradiation 4Gy-irradiated mice brains (according to the model SVZ regeneration after a moderate dose of irradiation that we previously described (Daynac et al. 2013). We used this model of SVZ reconstitution to gain insights on the annotation and and temporal ordering of neural progenitors clusters.
Project description:Spermatogenesis is a recurring differentiation process that results in the production of male gametes within the testes. During this process, spermatogonial stem cells differentiate to form spermatocytes, which undergo two rounds of meiotic division to form haploid spermatids. Throughout spermiogenesis, round spermatids elongate to form mature sperm. To profile maturing germ cells during the first wave of spermatogenesis, we generated droplet-based single cell RNAseq from juvenile animals at post-natal day P5-P35 as well as adult animals. Cells were isolated from whole testes. Furthermore, to assay the robustness of the meiotic cell division process, we profiled germ cells of the trans-chromosomal mouse model (Tc1) that carries a copy of the human chromosome 21.
Project description:In this study, we investigated somatic mutations in T cells in patients with various hematological disorders. To analyze immune cell phenotypes with somatic mutations, we performed scRNA+TCRab sequencing from 9 patients with chronic GVHD and clonal expansions of CD4+ or CD8+ T cells based on T cell receptor sequencing. CD45+ PBMCs (lymphocytes and monocytes) were sorted with BD Influx cell sorter and subjected to sequencing with Chromium VDJ and Gene Expression platform (v1.1, 10X Genomics). Sequencing was performed with Novaseq 6000 (Illumina). The immune cell phenotypes were compared to healthy controls processed in the same laboratory (accession number E-MTAB-11170). Due to data privacy concerns, the raw sequencing data is in the European Genome-Phenome Archive (EGA) under accession code [xxxx] and can be requested through the EGA Data Access Committee.
Project description:In the study we show that a specific peripheral glial population, derived from boundary cap (BC) cells, constitutes a major source of mural cells for the developing vasculature. Using Cre-based reporter cell tracing and single cell transcriptomics, we show that BC cell derivatives migrate along the nerves and differentiate into pericytes and vascular smooth muscle cells in the skin. The switch from glial to mural molecular identity is initiated while the cells are still associated with nerves To further characterize this transition glial to vascular identity, we performed single cell transcriptomic analyses (scRNA-seq) on FACS-purified traced cells from dissociated E12.5 skin. Tomato-positive cells were isolated by FACS from embryonic skin at E12.5. Around 10,000 cells were loaded into one channel of the Chromium system using the V3 single cell reagent kit (10X Genomics) We analyzed 2527 single cell transcriptomes with a mean number of expressed genes per cell of 4,696. This study highlights the plasticity of BC derivatives and uncovers a novel, nerve-derived origin for skin mural cells.
Project description:Understanding dynamics of antigen specific B cell responses and link between BCR and B cell differentiation is crucial for our ability to direct immune responses and to generate memory and plasma cells responses to protective targets. Here we have infected mice with IAV-PR8 and sacrificed them at different days after infection (7-14-28). 2 naive mice were used as controls (spleen and lungs). From individual spleen, lungs and mediastinal lymph nodes we have sorted Hemagglutinin (HA)-specific IgD- B cells and single cell sequenced RNA and BCR. We identify several known and novel B cell subpopulations forming after infection and find organ-specific difference that persist over the course of the response. We found important transcriptional differences between memory cells in lungs and immune organs and describe organ-restricted clonal expansion. Strikingly, by combining BCR mutational analysis, monoclonal antibody expression and affinity measurements we found no differences between germinal center (GC)-derived memory and plasma cells, at odds with an affinity-based selection model. These finding provide the most comprehensive picture to date of organ specific antiviral B cell responses, differentiation, clonal-proliferation and dynamics.