College - Author 1

College of Engineering

Department - Author 1

Biomedical Engineering Department

Degree Name - Author 1

BS in Biomedical Engineering

Date

5-2025

Primary Advisor

Trevor Cardinal, College of Engineering, Biomedical Engineering Department

Abstract/Summary

Peripheral artery occlusive disease (PAOD) is a progressive vascular condition that affects over 200 million people worldwide and approximately 8 million in the United States. It is characterized by atherosclerotic narrowing of peripheral arteries, leading to reduced blood flow, ischemia, and, in severe cases, tissue loss and limb amputation. Although surgical revascularization is the gold standard, many patients—particularly those with comorbidities like diabetes— continue to experience poor outcomes, emphasizing the need for alternative therapies. One promising strategy is to enhance collateral arteriogenesis, the remodeling and enlargement of collateral arterioles that naturally bypass occluded vessels. Satellite cells (SCs), which give rise to myogenic precursor cells, such as myoblasts, promote vascular regeneration through cytokine secretion and stimulation of arteriogenesis. However, laborious myofiber isolations which often yield relatively low numbers of primary myoblasts pose an obstacle to conducting cell transplantation studies. In this study, we supplemented culture media with hepatocyte growth factor (HGF)—a known activator of quiescent SCs—that we hypothesized could enhance myoblast yield. Myofibers were isolated from the extensor digitorum longus (EDL) muscle of C57BL/6 mice, and cultures were treated with one of three conditions: 0.1µg∙mL-1 bFGF alone, 10ng∙mL-1 HGF, or a combination of both. Myoblast proliferation was assessed daily via ImageJ based counts and on Day 5 via automated cell counting. bFGF-treated cultures had significantly greater proliferation on Days 4 and 5 compared to both HGF alone and the combination treatment; although the combination group followed a similar proliferative trend to bFGF, it did not surpass bFGF alone in effectiveness, suggesting a lack of synergy or potential antagonism. Future studies could explore optimization of HGF dosing and timing, as well as assess myogenic phenotype using RT-qPCR for markers such as MyoD and Pax7. Refinements to quantification methods, including DNA-quantification assays, may also address limitations in manual cell counting.

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