Human Skeletal Muscle Niche Formation and Analysis with Spatial RNA Sequencing

Formation of the human skeletal muscle can be achieved through xenotransplant of human stem or progenitor cells into mice. Human cells, such as those derived from human pluripotent stem cells (hPSCs), are dissociated from in vitro culture conditions and injected into immune-compromised mice where human cells must form new myofibers and retain or replace the mouse muscle stem cell pool. Efforts to better understand niche interactions will lead to improved regenerative potential that could ameliorate a broad range of muscle diseases. Spatial RNA sequencing of xenografted tissues allows for precise transcriptomic profiling of human muscle stem and progenitor cells in relation to myofibers and their niche throughout the myogenic differentiation process. Herein, we describe the procedures of obtaining high yields of human xenografted transplants and compare the use of various spatial RNA sequencing platforms to uncover stem cell niche formation.