Researchers at the University of Toronto have developed a new miniaturized assay platform, mini-MEndR, designed to evaluate muscle stem cell-mediated repair in a more efficient and scalable manner. This novel platform, funded by CFREF “Medicine by Design”, represents a significant advancement in the field of regenerative medicine, offering the potential to accelerate the discovery of therapeutic targets for muscle repair and regeneration.
The study, led by Profs. Penney Gilbert and Alison McGuigan, and recently published in BMC Methods, introduces a 96-well format assay that significantly enhances the throughput of skeletal muscle repair studies. This innovation addresses several bottlenecks in the original MEndR assay, which have previously limited its capacity to assess molecular targets on a large scale.
In comparison to alternative platforms, key advantages this platform offers are scalability, efficiency, quality assurance, and predictive capabilities.
Scalability: The new platform incorporates thrombin pre-adsorption to cellulose scaffolds, enabling in-situ hydrogel gelation. This method simplifies the manufacturing process, making it feasible to produce myotube templates using robotic liquid handling systems.
Efficiency: The miniaturized format cuts reagent costs in half and reduces hands-on seeding time by approximately threefold. Additionally, the image analysis pipelines save 40 hours of labor, allowing researchers to focus on data interpretation and follow-up experiments.
Quality Assurance: By evaluating multiple commercially available human primary myoblast lines in both 2D and 3D cultures, the researchers established quality assurance metrics that standardize myotube template quality, ensuring consistent and reliable results.
Predictive Capability: The Mini-MEndR platform replicates key results from the original MEndR assay, validating its ability to predict in vivo outcomes. This includes enhanced muscle production and Pax7+ cell expansion in response to p38/β MAPK inhibition, a known modulator of muscle stem cell-mediated repair.
“The Mini-MEndR platform offers a robust and scalable method to investigate human and mouse skeletal muscle endogenous repair. It holds promise for accelerating the muscle repair discovery pipeline, potentially leading to new therapeutic approaches for conditions such as aging-related muscle degeneration and Duchenne muscular dystrophy.” Said BME graduate student Nitya Gulati, one of the co-first authors of the study.
The research team aims to further integrate the mini-MEndR platform with high-content imaging systems and automated liquid handling robots to support large-scale screening efforts. Additionally, ongoing efforts will focus on reducing tissue replicate variability and demonstrating the platform’s application to disease modeling.