Meat and Medicine: Regenerative programming of animal and human stem cells for engineered skeletal muscle

Background: Pluripotent (PSC) and multipotent (such as mesenchymal; MSCs) stem cells hold great promise for the treatment of human skeletal muscle trauma and diseases, but also for the generation of new approaches to generate tissue for food consumption. However, it remains challenging to convert PSCs or MSCs to skeletal muscle cells, and the mechanisms by which the master regulatory transcription factors, promotes muscle stem (satellite) cell identity are not yet fully understood. However it is clear that viral or transgenic expression of transcription factors (TFs) such as Pax7 or MyoD can effectively induce the muscle program in these cell types.
Approach: We have developed a non-viral method to deliver genes to tissues, cells or stem cells. This will be the focus of the PhD, to deliver TF genes to stem cells in order to mediate there programming into myogenic precursors and to expand and use those cells to generate tissue of regenerative grafts and as meat-substitutes. You will use PSC and MSC lines as proof of concept, employing iPSCs from pig, sheep and cow to generate species specific muscle tissue for food applications. For clinical use you will employ human iPSCs, MSCs and bone marrow and liposuction aspirates to program these tissue stem cells into muscle for with cell-therapies or for tissue engineering and implant applications. The genes, duration of expression, if serial dosages are required and the culture conditions will require optimisation and assessment. The seeding of scaffolds with generated progenitors or directly programming in 3D will be needed to generate tissue which will also be studied. Interestingly we have electrical stimulation set ups that can be used to physiologically test
contraction of engineered muscle and there is potential to move to in vivo testing depending on the project with collaborators at the RCSI (Dublin, Ireland) and JHU (Baltimore, USA).

Location: The project will be carried out in the new state-of-the-art Biodiscovery institute at University Park (James Dixon) and the School of Biosciences at Sutton Bonington Campus (Ramiro Alberio). The use of human and animal cells of this application will be transformative and is likely to have significant commercial value and interaction with stakeholders. Presently the MoD (through the DSTL) are funding bone and fat reconstructive technologies using gene delivery in the Dixon group and this PhD could add impact and synergy to these projects reciprocally.

Techniques and opportunities: The project will use cell/stem cell culture, QPCR, immunolabelling, flow cytometry, single cell RNAseq, vector design and construction, gene delivery, scaffolds, tissue engineering, electrical stimulation, in vivo translation.
References to learn more:
1. Lilja et al. Pax7 remodels the chromatin landscape in skeletal muscle stem cells. PLoS ONE 12(4):e0176190.
2. Darabi R and Perlingueiro R (2016). Derivation of Skeletal Myogenic Precursors from Human Pluripotent Stem Cells Using Conditional Expression of PAX7. Methods in Mol Biol 1357:423-439