Project description:We have performed skeletal muscle organoid (SMO) cultures from three DMD patient-derived iPSC lines and one healthy iPSC control line. scRNA sequencing has been performed to specifically evaluate the myogenic progenitor / satellite cell populations of DMD versus healthy control SMOs.
Project description:The experiment aimed to validate the gene network regulating cell fate decisions during the myogenic differentiation of iPS cells from a DMD patient. DMD iPSCs were transfected with specific siRNA at Day 7 of the differentiation protocol and collected either at Day 9 or Day 14. As a control, we used a non-specific siRNA. Non transfected DMD cells were also included, together with the healthy control cell line used in our previous experiments.
Project description:Time Course in vitro Differentiation of Myogenic Primary Myoblast into Myotubes for Ontario Genome Project 2004-05. The Differentiation is leaded by removing the Proliferation Medium (Ham's F10 Medium + growth factors) and feeding with Differentiation Medium (DMEM hg + 2.5% Horse serum). The expected data are: % Viability of cells; Total RNA extraction; BioAnalysis and MicroArray of Undifferentiated and Differentiated cells. Time course: 0 hr, 6 hr, 12 hr, 18 hr, 24 hr, 36 hr, 48 hr, 3 days, 4 days, 7 days. Keywords: other
Project description:Myogenic Primary Myoblast Time Course in vitro Differentiation into Myotubes for Ontario Genome Project 2004-05. The Differentiation is leaded by removing the Proliferation Medium (Ham's F10 Medium + growth factors) and feeding with Differentiation Medium (DMEM hg + 2.5% Horse serum). The expected data are: % Viability of cells; Total RNA extraction; BioAnalysis and MicroArray of Undifferentiated and Differentiated cells. Time course: 0 hr, 6 hr, 12 hr, 18 hr, 24 hr, 36 hr, 48 hr, 3 days, 4 days, 7 days. Note that the Triplicate1 sample = MyoD-/-1999; Triplicate 2 sample = MyoD-/-p6; Triplicate 3 sample = MyoD-/-p10. Keywords: other
Project description:We previously demonstrated that iPSCs derived from a DMD patient and subjected to myogenic differentiation acquire a distinct transcriptomic profile from healthy controls. We observed a branched trajectory with an unbalanced distribution of Healthy and DMD cells on the two daughter branches. While each branch contained 86% of cells from a single line (either Healthy or DMD), we still observed 14% of cells from the other genetic background. This indicates that the choice between the two genetic states is probabilistic and does not strictly depend on the presence of the DMD mutation. In the present study, we hypothesize that cell fate choice at the bifurcation point is governed by a GRN. To gain insight into the molecular events directly downstream of the mutation, we generated more resolutive scRNA-Seq data around the bifurcation point. We subjected WT and mutant iPS cells to directed myogenic differentiation and we used Cellular Indexing of Transcriptomes and Epitopes by sequencing (CITE-Seq) to barcode cells collected at 8 time points spanning the bifurcation window between Day 5 and Day 14.
Project description:3 or 4 healthy cells - 3 DMD cells - 7 time points (tissue-derived myoblasts, tissue-derived myotubes, hiPSCs, Day 3 of differentiation, Day 10 of differentiation, Day 17 of differentiation, Day 25 of differentiation).
Project description:The goal of this study was to examine changes to gene expression induced by IL-13 treatment of air-liquid interface cultures of healthy primary human airway epithelia over time.