1. 3D model of human skeletal muscle egineered from
iPS cell-derived myogenic cells and biomaterials
Magdalena Plotczyk, Sara Martina Maffioletti, Dr Francesco Saverio Tedesco
Department of Cell and Developmental Biology, University College London
iPSCs and tissue engineering
Induced pluripotent stem cells (iPSCs) are somatic cells genetically
reprogrammed to stem cell-like state that possess an ability to differentiate
into various cell types of the body.
Patient-specific iPSCs can be cultured in vitro to generate progenitors and
differentiate to form 3D structures resembling tissues and organs -
organoids.
Magdalena Plotczyk
MSc Nanotechnology & Regenerative Medicine
University College London
magdalena.plotczyk.12@ucl.ac.uk
In order to improve cell culture conditions in 3D constructs, I investigated
the effect of an anti-apoptotic agent, ROCK inhibitor (ROCKi), on cell
viability and myogenic differentiation in 3D-cultured cells.
Fig. 2 The effect of ROCKi on cell viability in
3D. (A) Whole mount staining for MyHC on
HEDEMs differentiated for 10 days in 3D culture.
Nuclei counterstained with Hoechst. Scale bar
100μm. (B) Number of nuclei per mm3
quantified
based on merged confocal images. *P≤0.05,
unpaired t-test.
-ROCKi+ROCKi
MyHC, Hoechst
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Lancaster et al, 2013
Eiraku et al, 2011
iPSC-derived myogenic cells
In my project, I utilised human iPSC-derived myogenic cells to generate
3D skeletal muscle model.
iPSCs obtained from dystrophic patients were genetically corrected and
specified to become muscle progenitors.
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Skeletal muscle engineering platfom
Effect of ROCKi on cell viability in 3D constructs
Clinical and therapeutic applications
RESULTS
ROCK inhibitor can reduce apoptosis and increase cell density in 3D
culture of myogenic progenitors.
➔ 3D disease modelling
➔ personalised drug screening platform
➔ source of tissue replacement
➔ modelling myogenesis
The study was based on a muscle engineering platform combining
myogenic progenitors with a biomaterial, which was originally designed for
the culture of engineered heart tissue.
(Hansen et al, 2010)
Figure 1. Hydrogel
setup. (A) Freshly
polymerised gels (B)
from the bottom view
(C) and top view.
Scale bar 1cm.
Pictures taken by Sara
Martina Maffioletti
(unpublished).
A
B
C
Muscular dystrophies
Muscular dystrophies are inherited muscle disorders that cause
progressive muscle wasting and severely impaired mobility.
Duchenne Muscular Dystrophy is caused by mutations in dystrophin gene
encoding a protein responsible for cell membrane integrity. Breakdown of
the membrane leads to repeated cycles of degeneration and
regeneration, resulting in stem cell pool exhaustion and inability to repair
tissue.
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