In-vitro culture longevity has long been a concern for drug testing and disease modeling. The dynamic nature of certain cells, like skeletal muscle, contributes to their early peeling off the surface substrate and limits the system’s ability to carry out long term studies of any sort. To this end, we show an increase in the adhesion period of human-derived skeletal muscle cells up to 80 days. After watching muscle cultures peel as a sheet of myotubes at once, our hypothesis was to investigate ways to prolong cell-surface interactions. By means of regulating extracellular matrix (ECM) dynamics, specifically through inhibition of matrix metalloproteinases (MMPs) that work to digest components of the ECM, we show that the cells can stay adhered for longer periods of time with functional readouts. Functional testing showed that inhibitor treated cells displayed significantly reduced fatigue index and a higher fidelity than untreated cultures. Culture treatment with an MMP inducer showed premature peeling of the myotubes. Gel zymography data served as a proof of principle where cells treated with an inhibitor showed minimally active MMPs, while inducer-treated cells showed high MMP activity. All of this data supports the idea that regulating ECM dynamics through MMP inhibition can serve to maximize in-vitro myotube longevity. The result yields a possibility of more robust in-vitro systems with downstream potential for more accurate long-term toxicology assessments and disease modeling.
Bachelor of Science (B.S.)
College of Sciences
Length of Campus-only Access
Barakat, Natali, "Inhibition of Metalloproteinases Extends Longevity and Function of in-vitro Aged Human iPSC-Skeletal Muscle" (2022). Honors Undergraduate Theses. 1446.