Wragg_biot.201900106.pdf (3.32 MB)
Download file

Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system

Download (3.32 MB)
journal contribution
posted on 10.09.2019, 15:24 by Nick Wragg, Diogo Mosqueira, Lia Blokpeol‐Ferreras, Andrew CapelAndrew Capel, Darren J Player, Neil MartinNeil Martin, Yang LiuYang Liu, Mark LewisMark Lewis
In vitro three‐dimensional (3D) tissue engineered (TE) structures have been shown to better represent in vivo tissue morphology and biochemical pathways than monolayer culture, and are less ethically questionable than animal models. However, to create systems with even greater relevance, multiple integrated tissue systems should be recreated in vitro. In the present study, the effects and conditions most suitable for the co‐culture of TE skeletal muscle and bone were investigated. High‐glucose Dulbecco's Modified Eagle Medium (HG‐DMEM) supplemented with 20% foetal bovine serum (FBS) followed by HG‐DMEM with 2% horse serum was found to enable proliferation of both C2C12 muscle precursor cells and TE85 human osteosarcoma cells, fusion of C2C12s into myotubes, as well as an up‐regulation of RUNX2/CBFa1 in TE85s. Myotube formation was also evident within indirect contact monolayer cultures. Finally, in 3D co‐cultures, TE85 collagen/hydroxyapatite constructs had significantly greater expression of RUNX2/CBFa1 and osteocalcin/BGLAP in the presence of collagen‐based C2C12 skeletal muscle constructs; however, fusion within these constructs appeared reduced. This work demonstrates the first report of the simultaneous co‐culture and differentiation of 3D TE skeletal muscle and bone, and represents a significant step towards a full in vitro 3D musculoskeletal junction model.

Funding

Doctoral Training Centre of Regenerative Medicine, (EPSRC, UK)

History

School

  • Mechanical, Electrical and Manufacturing Engineering
  • Sport, Exercise and Health Sciences

Published in

Biotechnology Journal

Volume

15

Issue

1

Pages

1900106

Publisher

Wiley

Version

VoR (Version of Record)

Rights holder

© The Authors

Publisher statement

This is an Open Access Article. It is published by Wiley under the Creative Commons Attribution 4.0 Unported Licence (CC BY). Full details of this licence are available at: http://creativecommons.org/licenses/by/4.0/

Acceptance date

19/07/2019

Publication date

2019-09-20

Copyright date

2019

ISSN

1860-6768

eISSN

1860-7314

Language

en

Depositor

Dr Nick Wragg

Article number

1900106