Decellularized fetal skeletal muscle as an in vitro system to study a congenital muscular dystrophy

• Autor(a) principal: Soares, Ana Rita Gomes
• Data de Publicação: 2023
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10362/160290
• topic: Laminin-α2-congenital muscular dystrophy; skeletal muscle; extracellular matrix; decellularization; myogenesis; Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias

Laminin-α2-congenital muscular dystrophy (LAMA2-CMD) is caused by mutations in the LAMA2 gene, which encodes for the α2 chain of laminin-211, a glycoprotein present in extra-cellular matrix (ECM). Studies in our laboratory using a mouse model of the disease (dyW mice) have shown defects in fetal myogenesis. To better understand the impact of both extracellular environment and the cellular niche dur-ing the onset of LAMA2-CMD, wildtype C2C12 myoblast (C2C12-WT) and Lama2-null my-oblasts (C2C12-KO) were cultured in flasks and in decellularized fetal skeletal muscle obtained from embryonic day 18.5 healthy and dyW-/- fetuses. Our findings suggest that the absence of laminin-α2 results in alterations in the expression levels of genes that encode for other ECM proteins. Additionally, our results also show that proliferation and differentiation are also affected. Indeed, although C2C12-KO cells cultured in decellularized matrices showed a higher expression level of myogenin, a differentiation marker, and a higher tendency to formed aligned cells in matrices, only C2C12-WT cells were able to fuse and form multinucleated myotubes. The use of this novel 3D method allowed to reach a better understanding of the precise effects of laminin-α2 deficiency, which will be a requisite for the development of new therapies in the future.