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Developing and optimising defined tissue culture surfaces for cellular differentiation of muscle.

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  • Full or part time
    Prof M Peckham
  • Application Deadline
    No more applications being accepted
  • Funded PhD Project (European/UK Students Only)
    Funded PhD Project (European/UK Students Only)

Project Description

The study of cardiac and skeletal muscle in culture has the drawback that most commonly used approaches result in poorly differentiated cells, with a great deal of variability in their level of differentiation. This presents significant problems in attempts to quantify effects of disease mutations on sarcomere formation and organisation (myofibrilogenesis). Our goal is to improve in vitro muscle differentiation (cardiac and skeletal) using novel surfaces, developed in collaboration with Orla Technologies. Our recent collaboration with Orla Technologies, was recently published (Parker et al., 2016, Cytotechnology, epub Aug. 2016). In this successful collaboration, we used bio-active peptides derived from extracellular matrix proteins, engineered into the variable loops of the outer membrane protein A (OmpA). The purified protein is then applied stereospecifically onto a gold-coated surface via a cysteine residue in OmpA, generating an oriented monolayer, with peptides all facing upwards, accessible to the cells in a functional conformation. For skeletal muscle myotubes, we tested various combinations of laminin α2 peptides together with FGF, and obtained conditions in which myotubes differentiated on glass coverslips, with much higher levels of fusion and sarcomere formation, compared to controls.

This PhD studentship will now explore and develop this technology further in collaboration with Orla Technologies.

Our overall aims are to develop this approach for
• Patterned 2D surfaces. To better organise myotube formation, and obtain more uniform levels of differentiation across cells.
• Surfaces with variable stiffness. To find conditions that further improve myotube differentiation, known to be influenced by the stiffness of the growing surface and combine this with Orla Peptides. This approach can also be used for patterning (e.g. see Vignaud et al., ’Polyacrylamide Hydrogel Micropatterning’, Methods in Cell Biology, 120, p93: 2014)
• 3D culture. We will develop the techniques to generate ’tubes’ in which to grow myoblasts, that fuse into muscle cells. This approach may also be suitable for live cell imaging of muscle differentiation using light sheet.
• Differentiation of neonatal cardiomyocytes, including those differentiated from iPS cells suitable for the study of heart disease.

The methodology we will use will develop our current work with specific laminin, and other ECM peptides, developed with Orla allowing us to engineer and test new peptides if required.

This is a CASE studentship, and the student will have the opportunity to spend time at Orla Technologies in Newcastle as part of the studentship.

Funding Notes

Fully funded BBSRC iCASE studentship, providing UK/EU level fees plus a stipend (£14,296) for 4 years. EU candidates must have resided in the UK for three years prior to the start of the PhD to be eligible for full support; without evidence of residency the studentship will only provide fees and no stipend. Non-UK/EU candidates are not eligible. Candidates should have or be expecting a 2.1 or above at undergraduate level in a relevant subject. If English is not your first language you will be required to meet our English language requirements. The start date will be Oct 2017.


Azioune et al., (2010) Protein Micropatterns: A Direct Printing Protocol Using Deep UVs. Methods in Cell Biology, 97. p133
Vignaud et al., (2014) Polyacrylamide Hydrogel Micropatterning. Methods in Cell Biology. 120. p93
Parker et al., (2016) Promoting differentiation of cultured myoblasts using biomimetic surfaces that present alpha-laminin-2 peptides. Cytotechnology 68(5):2159-69
Clark et al., (1997) Preferential adhesion to and survival on patterned laminin organizes myogenesis in vitro. Exp. Cell. Res. 230: 275-83

How good is research at University of Leeds in Biological Sciences?

FTE Category A staff submitted: 60.90

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