RT Journal Article
SR Electronic
T1 Establishment and characterisation of primary skeletal muscle cell cultures from patients with advanced Chronic Kidney Disease
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.11.16.384263
DO 10.1101/2020.11.16.384263
A1 Luke A Baker
A1 Tom F O’Sullivan
A1 Kate A Robinson
A1 Zoe Redshaw
A1 Matthew Graham-Brown
A1 Robert U Ashford
A1 Alice C Smith
A1 Andrew Philp
A1 Emma L Watson
YR 2020
UL http://biorxiv.org/content/early/2020/11/16/2020.11.16.384263.abstract
AB Skeletal muscle wasting and dysfunction is a common characteristic of non-dialysis dependent chronic kidney disease (NDD-CKD). The mechanisms by which this occurs are not clearly understood and one reason for this is a lack of well controlled in-vitro methodologies to simulate NDD-CKD induced muscle wasting for mechanistic investigation at the cellular level. Here we sought to conduct the initial investigations into developing a CKD-induced skeletal muscle model for use as a mechanistic analysis tool as well as a test bed for potential novel therapeutics in this population. Human derived muscle cells (HDMCs) were isolated from n=5 NDD-CKD patients and n=3 matched healthy controls (HC) and taken through proliferation and differentiation phases in cell culture. Upon comparison of the 2 donor types, significantly greater mRNA expression of myogenic markers was noted in the NDD-CKD cultures in comparison to HC cultures, which was carried through to greater mRNA expression of myosin heavy chains (MyHCs) post differentiation. However, this was not carried over to protein expression where Pax7 and MyoD were seen to be expressed to a greater extent in HC cultures. mRNA expression markers of protein degradation were noted to be elevated in NDD-CKD cultures in comparison to HC cultures. In light of our findings, future work should seek to investigate the role of the ‘CKD environment’ as well as mechanisms implicated in transcription regulation to further advance the current model development as well as the mechanistic understanding of skeletal muscle wasting in CKD.Competing Interest StatementThe authors have declared no competing interest.