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Do factors secreted from synovial fibroblasts affect the differentiation of C2C12 cells?

McCabe, Emma Louise (2011)
M.Res. thesis, University of Birmingham.

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Abstract

The Wnt signalling pathway plays a key role within muscle differentiation. Wnt3a, Wnt5a, and DKK1 all have pivotal roles within this pathway. It’s hypothesised that due to their role within the Wnt pathway, Wnt3a, Wnt5a, and DKK1 will affect the differentiation of muscle, demonstrated by the murine C2C12 cell line.
Differentiation of the C2C12 cells was induced by adding DMEM differentiation media at day 0. Treatments (Wnt3a, Wnt5a, DKK1, control conditioned media, TNF-α conditioned media, and dexamethasone conditioned media) were added day 0, and mRNA levels of differentiation markers, MyoD, myogenin, α-actinin, and 11βHSD1 were measured using RT-PCR at days 1, 3 and 6. Wnt3a and control conditioned media gave no significant change in differentiation. Wnt5a, DKK1, TNF-α conditioned media and dexamethasone conditioned media gave significant decreases in differentiation. DKK1 inhibitor was tested on cells treated with TNF-α conditioned media, resulting in the decrease in the differentiation no longer being significant. 11βHSD1 enzyme activity assays were carried out to test Wnt3a, Wnt5a, DKK1, and DKK1 inhibitor effects, the results followed the trend of the mRNA data, however were not statistically significant. The results suggest that factors secreted from synovial fibroblasts during inflammation affect muscle differentiation.

Type of Work:M.Res. thesis.
Supervisor(s):Hardy, R.
School/Faculty:Colleges (2008 onwards) > College of Medical & Dental Sciences
Department:School of Clinical and Experimental Medicine, Centre for Endocrinology, Diabetes, and Metabolism
Additional Information:

Part 1 of a 2-part thesis. Part 2 is at http://etheses.bham.ac.uk/2858/

Subjects:QH301 Biology
Institution:University of Birmingham
ID Code:2857
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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