Project description:In order to assess the descendants of hypertrophic chondrocytes, we utilized Collagen10-Cre;Rosa26-tdTomato mouse total bone isolated at e16.5 by Collagenase II digestion after mechanical digestion and soft tissue removal. After sequencing and downstream analysis using Seurat, we observed clusters of cells with gene profiles matching classically defined chondrocytes, skeletal stem and progenitor cells (SSPCs), and osteoblasts. Trajectory analysis reveals that the SSPCs lie intermediate to the transition of chondrocyte to osteoblast. We conclude that hypertrophic chondrocytes dedifferentiate to this progenitor stage before further differentiation.
Project description:In order to assess the descendants of hypertrophic chondrocytes, we utilized Collagen10-Cre;Rosa26-tdTomato mouse bone marrow harvested at 2 months of age by centrifugation and light Collagenase II digestion. After sequencing and downstream analysis using Seurat, we observed clusters of cells with gene profiles matching classically defined skeletal stem and progenitor cells as well as CXCL12 abundant reticular (CAR) cells. These cells appear to be upstream of both osteoblasts and adipocytes. We conclude that hypertrophic chondrocytes dedifferentiate to this progentior stage before further differentiation.
Project description:To investigate the effects of Dnmt1 Knockout in Sst expressing interneurons on celltype distribution in the embryonic brain, Sst-Cre/tdTomato/Dnmt1 loxp2 knockout mice and Sst-Cre/tdTomato control mice were analysed. 10 µm coronal sections of E16.5 brains were prepared for the Merscope platform using the commercially available Pan Neuro panel (Vizgen), covering 500 distinct transcripts.
Project description:In order to assess the descendants of lateral plate mesoderm within the muscle interstitium, we utilized Prrx1Cre;Rosa26-tdTomato P21 tdTomato+ FACS sorted muscle interstitial cells
Project description:Vascular smooth muscle cells (VSMCs) in the ascending aorta and aortic arch arise from two distinct embryonic origins: the second heart field (SHF) and the cardiac neural crest (CNC). Whether these populations exhibit sex-dependent differences remains unknown. Here, we used Wnt1-Cre;Rosa26-mTmG mice to genetically label CNC-derived VSMCs with GFP, while non-CNC-derived cells expressed tdTomato. The ascending aorta and aortic arch were dissected from male and female mice, and single-cell suspensions were generated by enzymatic digestion. Cells were then sorted by fluorescence-activated cell sorting (FACS) based on GFP or tdTomato expression. RNA-expression profiles for each group were obtained using the Smart-seq platform. Thi study revealed the differential expression of VSMCs from different embryonic origin in both sexes.
Project description:Comparison of aP2-Cre and Cdh5-Cre tdTomato labaled cells from hindlimb and SCM muscles shows they mark the same endothelial cell populations