Poster Presentation 28th Annual Lorne Proteomics Symposium 2023

Adipose derived stem cells spontaneously express neural markers when grown in a PEG-based 3D matrix (#120)

Neus Gomila Pelegri 1 2 , Bruce K Milthorpe 1 , Amy Bottomley 3 , Catherine A Gorrie 2 , Matthew P Padula 4 , Jerran Santos 1
  1. Advanced Tissue Engineering and Stem Cell Biology Group, School of Life Sciences, University Of Technology Sydney, Sydney, NSW, Australia
  2. Neural Injury Research Unit, School of life Sciences, University of Technology Sydney, Sydney, NSW, Australia
  3. Microbial Imaging Facility (MIF) AIMI, , Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia
  4. School of life sciences, University of Technology Sydney, Sydney, NSW, Australia

Neurological diseases are among the leading causes of disability and death worldwide, are economically costly, reduce the quality of life and are difficult to treat as they are poorly understood. Tissue engineering offers potential treatment avenues for these conditions; however, the development of biologically accurate models of brain tissues remains challenging. Adipose-derived stem cells (ADSCs) are a multipotent cell population of interest for the creation of neural tissue models given their neurogenic potential and availability. While progress has been made in differentiating ADSCs into neural cells, with chemical inductions in 2D environments being the most common method, differentiation in 3D environments that are more representative of the in vivo physiological conditions of the nervous system may be beneficial. One of the ways this can be achieved is through modulating the 3D matrix composition and stiffness that the cells are grown in. To address this, we cultured ADSCs for 14 days in a 1.1kPa PEG-based hydrogel matrix containing adhesion peptides YIGSR and RGD. To assess the effects of the matrix on the cells we assessed morphology using live cell imaging, cell viability using Alamar blue viability assay, proteome changes using untargeted proteomics and spontaneous neural differentiation using immunocytochemistry of neural markers. The cells grown in the 3D matrix were compared to those grown in 2D conditions. Results showed that the cells continued to proliferate over the 14-day period and presented a different morphology to the 2D cultures, with the cells elongating and aligning with one another. Additionally, immunocytochemistry results showed that cells started to spontaneously express the CNPase oligodendrocytic marker in the 3D cultures. The proteome analysis revealed that 86 proteins changed in abundance by >1.5 fold. Of particular interest, Gamma-enolase-2 protein, a well-known neuronal marker, increased by 4.08 fold. These findings are indicative that ADSCs will start to spontaneously increase neural marker expression when grown in an environment that mimics the physical environment of the nervous system.