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Optimization of Satellite Cell Culture Through Biomaterials

  • Sadegh Davoudi
  • Penney M. GilbertEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1556)

Abstract

Hydrogels, a type of biomaterial, are an invaluable part of biomedical research as they are highly hydrated and properties such as elasticity, porosity, and ligand density can be tuned to desired values. Recently, culture substrate stiffness was found to be an important regulator of muscle stem cell self-renewal. Polyethylene glycol (PEG), a synthetic polymer, can be fabricated into hydrogels that match the softness of skeletal muscle tissue, thereby providing a culture surface that is optimal for maintaining muscle stem cell self-renewal potential ex vivo. In this Chapter, we describe a method to produce flat PEG hydrogels across a range of stiffnesses, including a formulation that matches the bulk stiffness of healthy skeletal muscle (12 kPa), while maintaining a constant ligand density. Since PEG is inert to protein adsorption, the steps required to surface functionalize the hydrogel with an adhesive interface (e.g., laminin) are also described.

Key words

Muscle stem cell Hydrogel Polyethylene glycol Substrate stiffness Culture substrate Self-renewal 

Notes

Acknowledgment

The Natural Sciences and Engineering Research Council and the Canadian Institutes of Health Research supported this work.

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Copyright information

© Springer Science+Business Media LLC 2017

Open Access This chapter is distributed under the terms of the Creative Commons Attribution Noncommercial License, which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Authors and Affiliations

  1. 1.Institute of Biomaterials and Biomedical EngineeringUniversity of TorontoTorontoCanada
  2. 2.Donnelly Centre for Cellular and Biomolecular ResearchUniversity of TorontoTorontoCanada

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