Development of a Tissue Engineered Skeletal Muscle Construct Containing Lipids as a Model to Study Diabetes
The goal of this research project was to develop and characterize an in vitro skeletal muscle construct that could be used as a model to study diabetes. To reach this goal the project has been subdivided into two parts. Firstly, the research was focused on identifying the culture conditions that can promote satellite cell (SC) adipogenic differentiation by using fibrin gels as basic model scaffold. qPCR analysis of key angiogenic and myogenic factors, as well as staining of lipid content, was carried out to evaluate the differentiation of SCs. The second part of this thesis was aimed at developing a functional tissue-engineered skeletal muscle construct. Fibrin scaffolds containing SCs from both lean or diabetic rat skeletal muscle tissues were fabricated in pre-fabricated molds containing pins that created a passive tension within the constructs. Because of this tension the cells were able to grow and spread unidirectionally, forming myogenic structures similar to those found in skeletal muscle tissue. The constructs were tested for their mechanical properties and their ability to promote glucose uptake upon insulin stimulation, relative to constructs containing cells taken from healthy animals. The diabetic constructs had decreased mechanical properties, reduced glucose intake, and higher expression of adipogenic genes. Overall, the engineered skeletal muscle tissue constructs that contained lipids could be used to study diabetes and may be helpful in identifying the biological cues that could reverse fat accumulation in skeletal muscle.