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

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 in...

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Main Authors:	Nick Wragg, Diogo Mosqueira, Lia Blokpeol‐Ferreras, Andrew Capel, Darren J Player, Neil Martin, Yang Liu, Mark Lewis
Format:	Default Article
Published:	2019
Subjects:	Bone Co‐culture Medium compatibility Skeletal muscle Tissue engineering
Online Access:	https://hdl.handle.net/2134/9778661.v1
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id	rr-article-9778661
record_format	Figshare
spelling	rr-article-97786612019-09-20T00:00:00Z Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system Nick Wragg (1260240) Diogo Mosqueira (1830001) Lia Blokpeol‐Ferreras (7353083) Andrew Capel (1258833) Darren J Player (7256603) Neil Martin (1258602) Yang Liu (1257603) Mark Lewis (1255254) Bone Co‐culture Medium compatibility Skeletal muscle Tissue engineering 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. 2019-09-20T00:00:00Z Text Journal contribution 2134/9778661.v1 https://figshare.com/articles/journal_contribution/Development_of_a_3D_tissue-engineered_skeletal_muscle_and_bone_co_culture_system/9778661 CC BY 4.0
institution	Loughborough University
collection	Figshare
topic	Bone Co‐culture Medium compatibility Skeletal muscle Tissue engineering
spellingShingle	Bone Co‐culture Medium compatibility Skeletal muscle Tissue engineering Nick Wragg Diogo Mosqueira Lia Blokpeol‐Ferreras Andrew Capel Darren J Player Neil Martin Yang Liu Mark Lewis Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
description	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.
format	Default Article
author	Nick Wragg Diogo Mosqueira Lia Blokpeol‐Ferreras Andrew Capel Darren J Player Neil Martin Yang Liu Mark Lewis
author_facet	Nick Wragg Diogo Mosqueira Lia Blokpeol‐Ferreras Andrew Capel Darren J Player Neil Martin Yang Liu Mark Lewis
author_sort	Nick Wragg (1260240)
title	Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
title_short	Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
title_full	Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
title_fullStr	Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
title_full_unstemmed	Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system
title_sort	development of a 3d tissue-engineered skeletal muscle and bone co‐culture system
publishDate	2019
url	https://hdl.handle.net/2134/9778661.v1
_version_	1799724570918256640

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

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