Project description:Transcriptional profiling of mouse kidney papilla cells comparing control GFP- cells with GFP+ labeled cells. Mouse kidney papillae were dissected. GFP- and GFP+ cell populations were isolated by FACS. The latter represent slow cycling label retaining cells (LRCs).
Project description:Different types of hair follicles can be found in the skin of mice. It is believed that the signals that control hair follicle differentiation arise from cells in a structure called the dermal papilla. Understanding the nature of those signals is of interest for the biology of the normal tissue. We have developed a technique for isolation of dermal cells by enzymatic digestion of intact skin. We have identified two subpopulations of cells that can be separated by FACS. The Sox2-positive CD133-positive cells are found exclusively in the dermal papillae of guard/awl/auchene hairs, while Sox2-negative, CD133-positive cells are found in the other hair follicle types. We compared these populations with unfractionated dermal cells. We isolated the following 3 populations of cells from the back skin of neonatal mice (P2) by Flow Cytometry: 1) GFP-CD133- Total dermal cells 2) GFP-CD133+ Dermal Papilla cells 3) GFP+CD133+ Dermal Papilla cells The yield is approximately 50,000 cells of each population.
Project description:Label retaining and non-retaining muscle stem cells from young and aged H2B-GFP+/-;rtTA+/- were profiled by single cell RNA-seq at two timepoints
Project description:Homeostatic hematopoietice stem cells (HSCs) with greater divisional history lose repopulating potential after very few cell divisions. Divisional history overrides both phenotype and immediate quiescence in determining functional activity. In GFP label retaining system GFP is progressively diluted when cells proceed through a cascade of divisions. We used a GFP label retaining system and performed microarray expression analyses to track the changes in the gene expression profile of bone marrow (BM) LSK cells that relates to divisional history during homeostasis.
Project description:Hierarchically organized tissues, such as hematopoietic systems, muscle, or skin harbor deeply quiescent stem cells which start proliferating in response to external insults. In contrast, it remains obscure whether similar quiescent cells exist in epithelia of digestive organs. Here we identified a deeply quiescent population in gastric corpus but not in other gastrointestinal organs after systematic examination of H2b-GFP label-retaining cells. The label-retaining cells in corpus epithelia belonged to a subpopulation of chief cells that were located near basal layers of corpus and did not overlap with Troyhigh, Lgr5high, or Misthigh cell population. The identified quiescent cells were marked with activation of Atf4 and unfolded protein response. External damages by indomethacin treatment triggered proliferation of the quiescent populations, indicating that chief cells of gastric corpus harbor deeply quiescent reserve cells with high levels of internal stress response activity.
Project description:This study was performed to compare transcriptomic changes in the heterogeneous mouse skin epidermal stem cells and mouse oral epithelial stem cells during homeostasis at young age. Slow-cycling stem cells (label retaining cells, LRCs), fast-cycling stem cells (non-label retaining cells, nLRCs) express unique gene signatures in both skin and oral tissues and have independent stem cell identities. Our data indicates that there are similarities and differences in gene expression patterns between LRC and non-LRC populations in different epithelial tissues, suggesting stem cell autonomous heterogeneity and environmental differences.
Project description:The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyse the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signalling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry. </br></br> The CE-MS assay is reported in the current study MTBLS633. </br> The General LC-MS assay is reported in MTBLS629. </br> The Lipidomic LC-MS assay is reported in MTBLS631. </br> The GC-MS assay is reported in MTBLS634. </br></br> Linked Studies: <a href='https://www.ebi.ac.uk/metabolights/MTBLS629' target='_blank'><span class='label label-success'>MTBLS629</span></a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS631' target='_blank'><span class='label label-success'>MTBLS631</span></a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS634' target='_blank'><span class='label label-success'>MTBLS634</span></a>
