Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of naive CD4 T cells. By sequencing this population in young and older donors, we describe the heterogeneity of Tscm CD4 cells based on differential engagement of Wnt pathways and inflammation.
Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of naive CD4 T cells. By sequencing this population in young and older donors, we describe the heterogeneity of Tscm CD4 cells based on differential engagement of Wnt pathways and inflammation.
Project description:Gene expression data of primary human naive and memory CD4+T lymphocytes purified from peripheral blood are generated to be analyzed in different ways such as for traditional searching of differentially expressed genes between the two cell subsets or in combination to in-silico data of microRNAs target prediction for microRNAs known to be characteristically expressed in the two cell subsets. Two cell subsets (cell types) FACS purified from peripheral blood of six samples/healthy donors (samples #3,5,6,7,026,065). Naive CD4+ T cells were extracted from all 6 samples and 6 biological replicas were obtained (3N, 5N, 6N, 7N, 026N, 065N), while memory CD4+T cells were extracted from 4 samples and 4 biological replicas were obtained (3m, 5m, 6m, 7m). Both naive and memory replicas from samples 3, 5, 6 and 7 were hybridized onto two beadsarrays each while those from samples 026 and 065 were hybridized on 1 beadsarray each (for a total of 18 beadsarrays used). For analysis purposes naive cells samples 3N, 5N, 6N and 7N can be considered paired with memory samples 3m, 5m, 6m and 7m respectively, since they are obtained from the same blood samples/healthy donors.
Project description:We detected and compared the miRNAs contained in Y-PRP-exos or O-PRP-exos. The results showed that most miRNAs (38) can be identified in both Y-PRP-exos and O-PRP-exos. In addition, hsa-let-7f-5p, hsa-miR-16-5p and hsa-miR-126-3p were top 3 enriched miRNAs in PRP-exos of all donors. These results indicated the heterogeneity of miRNAs in PRP-exos was small, regardless of age. However, the abundance of miRNAs in Y-PRP-exos and O-PRP-exos was quite different, especially in high-ranking miRNAs. The expression of hsa-miR-16-5p and has-let-7f-5p in O-PRP-exos in two old donors was only approximately 1/4-1/2 of that in Y-PRP-exos in two young donors
Project description:Mesenchymal stem/stromal cells (MSCs) with immunosuppressive properties are increasingly used in advanced cellular therapies. Since the clinical use of hMSCs demands sequential cell expansions, we studied the effect of cell doublings on the phospholipid profile as well as functionality of human bone marrow mesenchymal stem cells (hBMSCs). In addition to the structural role of phospholipids in cell membranes, they provide precursors for eicosanoids and other signalling lipids modulating cellular functions. The hBMSCs, harvested from young adult and old donors (n=5 for both), showed clear compositional changes during cultivation, seen at the level of lipid classes, lipid species and acyl chains. As the main finding at the lipid class level, the ratio of phosphatidylinositol to phosphatidylserine was increased towards the late passage samples. In the species profiles, arachidonic acid (AA) containing species of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) clearly accumulated, while the species containing monounsaturated fatty acids decreased. This was related with an increase of AA, a major n-6 polyunsaturated fatty acid (n-6PUFA), in the total fatty acid pool of the cells, which happened at the expense of n-3PUFAs, especially docosahexenoic acid (DHA). Using hBMSCs from four of the young adult donors and four of the old donors, we found that gene expression of several enzymes involved in fatty acid metabolism (such as FADS1, FADS2 and SCD) was altered. The expression of genes related to the regulation of cell cycle, senescence and immunomodulation were altered. Our findings suggest that multistep expansion of hBMSCs alters their fatty acid metabolism and membrane phospholipid composition, which affects lipid signalling and eventually the immune function of the cells. Cultured undifferentiated bone marrow-derived MSCs from old and young donors. Biological replicates: 4 old donors and 4 young donors, passages 4 and 8 from each.
Project description:FACS-sorted CR2+ and CR2- naive and memory CD4+ T cells from healthy donors, plus CR2+ naive CD4+ T cells activated using CD3/CD28 beads, were analyzed ex-vivo. Exome-enriched RNA sequencing was carried out for all subsets.
Project description:Here, we present genome-wide comparisons of chromatin accessibility from endogenous CD8+ T cells and transferred OT-1 CD8+ T cells in young vs. aged B16-OVA tumors using ATAC-seq. This ATAC-seq dataset includes three different intratumoral CD8+ T cell subsets, such as TProg: Slamf6+Tim-3-, TTerm: Slamf6-Tim-3+, and TTAD: Slamf6-Tim-3-, as well as naive CD8+ T cells from spleens.
Project description:This study comprises DNA methylation profiles of 9 directly converted human fibroblasts from young, adult and old donors into oligodendrocytes (dc-hiOL).
Project description:Mesenchymal stem cells were isolated from several donors of different age (young vs old), and total RNA was extracted. Samples were labeled with Cy3 and Cy5 dyes and hybridized in a looped design that allowed the calculation of differentially expressed miRNAs in MSCs isolated from healthy, aged individuals.