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Myogenesis pp 169-183 | Cite as

Coculture Method to Obtain Endothelial Networks Within Human Tissue-Engineered Skeletal Muscle

  • Dacha Gholobova
  • Melanie Gerard
  • Lisanne Terrie
  • Linda Desender
  • Janet Shansky
  • Herman Vandenburgh
  • Lieven ThorrezEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1889)

Abstract

Skeletal muscle tissue engineering aims at creating functional skeletal muscle in vitro. Human muscle organoids can be used for potential applications in regenerative medicine, but also as an in vitro model for myogenesis or myopathology. However, the thickness of constructs is limited due to passive diffusion of nutrients and oxygen. Introduction of a vascular network in vitro may solve this limitation. Here, we describe tissue engineering of in vitro skeletal muscle consisting of human aligned myofibers with interspersed endothelial networks. To create bio-artificial muscle (BAM), human muscle progenitor cells are cocultured with human umbilical vein endothelial cells (HUVECs) in a fibrin hydrogel. The cell-gel mix is cast into silicone molds with end attachment sites and cultured in endothelial growth medium (EGM-2) for 1 week. The passive forces generated in the contracted hydrogel align the myogenic cells parallel to the long axis of the contracted gel such that they fuse into aligned multinucleated myofibers. This results in the formation of a 2 cm long and ~1.5 mm tick human BAM construct with endothelial networks.

Key words

Coculture Tissue engineering Skeletal muscle Myoblasts Endothelial cells Bio-artificial 

Notes

Acknowledgments

This work was funded by AFM-Telethon trampoline grant 19802 and the Research Foundation-Flanders (FWO) grant 1529817N.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Dacha Gholobova
    • 1
  • Melanie Gerard
    • 1
  • Lisanne Terrie
    • 1
  • Linda Desender
    • 1
  • Janet Shansky
    • 2
  • Herman Vandenburgh
    • 2
  • Lieven Thorrez
    • 1
    Email author
  1. 1.Tissue Engineering Laboratory, Department of Development and RegenerationKU Leuven campus KulakKortrijkBelgium
  2. 2.Department of Pathology and Laboratory MedicineBrown UniversityProvidenceUSA

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