Human Skeletal Muscle Niche Formation and Analysis with Spatial RNA Sequencing

Area of Science:

• Regenerative Medicine
• Stem Cell Biology
• Muscle Physiology

Background:

• Human skeletal muscle formation can be achieved via xenotransplantation of human stem or progenitor cells into mice.
• Human cells, including human pluripotent stem cells (hPSCs), must form new myofibers and integrate with the mouse muscle stem cell pool post-transplantation.
• Understanding niche interactions is crucial for enhancing regenerative potential in muscle diseases.

Purpose of the Study:

• To describe procedures for obtaining high yields of human xenografted muscle transplants.
• To compare different spatial RNA sequencing platforms for analyzing xenografted tissues.
• To uncover stem cell niche formation during myogenic differentiation in vivo.

Main Methods:

• Xenotransplantation of human stem/progenitor cells into immune-compromised mice.
• Dissociation of cells from in vitro culture and injection into mouse muscle.
• Spatial RNA sequencing for transcriptomic profiling of human cells within the mouse niche.

Main Results:

• Successful high-yield xenografted transplant procedures were established.
• Spatial RNA sequencing provided precise transcriptomic data of human cells in relation to their niche.
• Insights into stem cell niche formation and myogenic differentiation were gained.

Conclusions:

• The study provides a framework for generating and analyzing human skeletal muscle xenografts.
• Spatial transcriptomics is a powerful tool for dissecting stem cell niche dynamics.
• This research contributes to understanding and potentially treating muscle diseases through improved regenerative strategies.