Identification and Analysis of Myogenic Progenitors In Vivo During Acute Skeletal Muscle Injury by High-Dimensional Single-Cell Mass Cytometry

Our research investigates the molecular mechanisms underlying stem cell dysfunction in aging and disease. Our goal is to understand how muscle stem and progenitor cells interact with other cells in the muscle microenvironment to achieve effective regeneration. We are also studying whether a barren cell communication impacts muscle stem cell function in aging and disease.

A major barrier to investigating cell-cell communication within the stem cell niche has been the lack of technologies that provide single cell resolution. Single cell mass cytometry allows simultaneous high throughput quantitative analysis on multiple cell types and molecular phenotypes in complex tissues. Our protocol enables purification and deep profiling of rare stem cells and the progeny during muscle regeneration using antibodies to cell surface markers and myogenic transcription factors.

Studying these cells helps us understand how muscle stem cell fates is controlled, and uncover the networks that drive cell state transitions doing effective regeneration. Measuring up to 50 cell surface and intracellular protein simultaneously at a single cell level is a valuable approach for investigating cellular function in muscle tissue. This technique can identify unique molecular signatures and reveal novel mechanisms of muscle stem cell dysfunction in aging and disease.

This protocol allows to investigate how cell state transition impact tissue regeneration in health and disease. The signature of activated stem cells we present here enable studies of stem cell quiescence. They were not previously possible.

We can now purify activated stem cells, CD98 High, CD44 High, based solely on cell surface marker, and study for the first time, how they return to quiescence.