Background: Muscle wasting in ageing (sarcopenia) is a significant public health concern, culminating in weakness, frailty and loss of independence. Despite several decades of research using model organisms, animals and two dimensional (2D) muscle cell cultures, there remains no cure or treatment for muscle wasting. Animal models and cells do not represent human muscle ageing and disease, while 2D models do not recapitulate the 3D structure of muscle tissue and lack the most essential functional characteristic, muscle contraction, because they do not have a neural connection. Thus, new sophisticated models are required to more efficiently trial novel drugs and compounds designed to enhance muscle growth and regeneration.
Aim: The main aim of this project is to 1) establish a 2D Human co-culture nerve-muscle model 2) to transfer the 2D model to 3D human co-culture platform. 3) To test phenotypic changes after exposure to drugs.
Methods: Human myoblasts donated by a 25 yr old male (C25) and an 83 yr old male (C83) were separately seeded with growth media (GM) and incubated over 24 hours at 37°C within 5% CO2. Thereafter, cells were co-cultured with differentiation media (DM) to investigate differentiation parameters. Human neural progenitor cells (NPCs) cells were differentiated from embryonic stem cells to be co-cultured with human muscle cells (SkMC). 3D human Myobundles were formed using cell- hydrogel mixture placed on laser cut Farbric frames within biomaterial moulds. Results: 1) No variations were noted between C25 and C83 when observing the fusion index, myoblast region as well as aspect ratio following 96 hours. 2) NPCs were successfully generated. 3) 3D human myobundle was successfully established.