Project description:To determine changes in pancreatic mesenchymal cells during embryonic development, we analyzed the transcriptome of these cells at three embryonic days: 13.5 ('L1'), 15.5 ('L2'), and 17.5 ('L3'). Pancreatic mesenchymal cells were isolated by FACS from Nkx3.2-Cre;LSL-YFP pancreatic tissues based on these cells fluorescent labeling. Two groups of mice were analyzed for each embryonic day ('P1', 'P2').
Project description:To identify proteins phosphorylated by Akt downstream of PI3K-mediated PDGFRalpha signaling, we immunoprecipitated Akt phosphorylation substrates from PDGF-AA-treated primary mouse embryonic palatal mesenchyme (MEPM) lysates and analyzed the peptides by nano LC-MS/MS.
Project description:The surrounding microenvironment plays a crucial role in pancreas formation and differentiation. Nevertheless, its cellular composition has not been analyzed in depth and whether different cell populations exist within the mesenchyme of the developing pancreas is an open question. Similarly, the molecular mechanisms by which the mesenchyme coordinates and integrates various signaling pathways to stimulate epithelial pancreatic progenitor proliferation and differentiation remain elusive. Here, we investigate the cell composition and molecular network in the pancreatic mesenchyme, which promote pancreas differentiation. Whole transcriptome analysis and functional experiments showed that PBX in the mesenchyme promotes endocrine differentiation non-cell autonomously, acting at two different levels in the epithelial-mesenchymal crosstalk: 1) the maintenance of BM integrity and ECM and 2) regulation of soluble molecules, including guidance cues. In conclusion, this study has identified distinct pancreatic mesenchymal niches and dissected their different roles during pancreatic development. This knowledge will be useful to improve current differentiation strategies based on ECM and soluble factors to generate β-cell equivalents from pluripotent cells for future cell replacement therapies of diabetes.
Project description:Identification of genes differentially expressed between epithelium and mesenchyme of embryonic lung. Embryonic lung are harvested from time pregnant mice
Project description:Mouse embryonic stem cells containing a Sox17-GFP construct were differentiated using growth factors (Activin A and Wnt3A) to definitive endoderm. Sox17-GFP(+) cells were sorted using fluorescence activated cell sorting and either used for total RNA harvest OR continued in culture in the presence of primary pancreatic mesenchymal cell lines. At the end of 6 serial passages on mesenchyme, the Sox17-GFP(+) cells were again sorted and the RNA was harvested for arrays. Samples were prepared as described in summary, with technical duplicates for each of the following 3 categories: 1. Unpassaged (P0) endoderm, 2. Endoderm passaged 6 times (P6) on mesenchyme 1, and 3. Endoderm passaged 6 times (P6) on mesenchyme 2.
Project description:Analysis of steady-state mRNA levels in embryonic mouse facial mesenchyme tissue at E10.5 from wild-type C57BL/6, Dp16, and interferon receptor dosage-normalized Dp16^2xIFNRs mice. This dataset is part of the Human Trisome Project - Trisomy 21 Model Atlas run by the Linda Crnic Institute for Down Syndrome at the University of Colorado Anschutz Medical Campus. http://www.trisome.org/
Project description:The lung mesenchyme plays important roles in lung development and is affected in many respiratory diseases, yet relatively little is known about the biology of lung mesenchymal progenitors. We sought to establish an induced pluripotent stem cell (iPSC)-based model to study lung mesenchyme development and epithelial-mesenchymal interactions. We generated a mouse iPSC line carrying a lung mesenchyme-specific reporter/tracer to establish a protocol for differentiation into lung mesenchymal progenitors. We derived lung mesenchyme from iPSCs both by directed differentiation via a lateral plate mesodermal progenitor state (induced lung mesenchyme, iLM), and by co-development during lung epithelial differentiation (co-developed lung mesenchyme, cLM). We found that directed differentiation via a lateral plate mesoderm progenitor was not only more efficient, but also yielded engineered lung mesenchymal cells that were more similar to primary lung mesenchyme from day 12.5 mouse embryos, as determined by single cell RNAseq. Our iPSC-derived population will provide an inexhaustible source of cells for studying lung development, modeling diseases, and developing therapeutics.