Project description:We performed single cell RNA sequencing analysis of human iPSC-derived skeletal muscle organoids at 12 week post differentiation stage.
Project description:We performed RNA sequencing analyses of human iPSC and iPSC-derived skeletal muscle organoids in different differentiation stages of 4wk, 8wk, and 16wk.
Project description:To begin to validate potential causal regulators of muscle function, we targeted genes containing novel skeletal muscle pQTLs and molecular/phenotypic associations, and performed a functional genomic screen in human skeletal muscle organoids (PMID: 30527761). We focused on proteins with negative associations to lean mass, grip strength or other metabolic associations, and generated a total of 27 individual rAAV6:shRNAs. Organoids were grown around contraction posts to monitor contractile force production during electrical stimulation, and transduced following differentiation and maturation to limit effects on the myogenic program (Figure 3A). Electrical stimulation was performed to induce either a tetanic contraction for assessment of maximum force production or stimulated with sustained lower frequency for assessment of fatigue. Following the protocol, organoids were analysed by proteomics which quantified 17/27 targets with 13 targets significantly reduced in abundance by rAAV6:shRNA.
Project description:This study aims to characterize the diversity of cell types in human skeletal muscle across age using two complementary technologies: single-cell and single-nucleus sequencing, which provide a comprehensive coverage of cell types in the muscle. We leveraged the aforementioned datasets to study change in cell type composition and gene expression between young (n= 8, approx. 20-40 yrs) and old (n = 9, approx. 60-80 yrs) adults, highlighting changes in the major skeletal muscle compartments, muscle satellite cells, myofiber and muscle microenvironment including stromal, immune and vascular cell types. Additionally, we generated a complementary mouse muscle aging dataset by profiling hindlimb muscles from young (n = 5, 3 months) versus old mice (n = 3, 19 months), using single-cell and single-nucleus sequencing for comparison.
Project description:Skeletal muscle accounts for the largest proportion of human body mass, on average, and is a key tissue in complex diseases and mobility. It is composed of several different cell and muscle fiber types. Here, we optimize single-nucleus ATAC-seq (snATAC-seq) to map skeletal muscle cell-specific chromatin accessibility landscapes in frozen human and rat samples, and single-nucleus RNA-seq (snRNA-seq) to map cell-specific transcriptomes in human. We additionally perform multi-omics profiling (gene expression and chromatin accessibility) on human and rat muscle samples.