Matrigel 3D bioprinting of contractile human skeletal muscle models recapitulating exercise and pharmacological responses

Angela Alave Reyes-Furrer,Sonia De Andrade,Nathalie Accart,Martin Steinmann,Dominic Bachmann,Andrew Dunbar,Epifania Bono,Martin Rausch,Hansjoerg Keller,Heidi Jeker,Markus Rimann

doi:10.1038/s42003-021-02691-0

Abstract

A key to enhance the low translatability of preclinical drug discovery are in vitro human three-dimensional (3D) microphysiological systems (MPS). Here, we show a new method for automated engineering of 3D human skeletal muscle models in microplates and functional compound screening to address the lack of muscle wasting disease medication. To this end, we adapted our recently described 24-well plate 3D bioprinting platform with a printhead cooling system to allow microvalve-based drop-on-demand printing of cell-laden Matrigel containing primary human muscle precursor cells. Mini skeletal muscle models develop within a week exhibiting contractile, striated myofibers aligned between two attachment posts. As an in vitro exercise model, repeated high impact stimulation of contractions for 3 h by a custom-made electrical pulse stimulation (EPS) system for 24-well plates induced interleukin-6 myokine expression and Akt hypertrophy pathway activation. Furthermore, the known muscle stimulators caffeine and Tirasemtiv acutely increase EPS-induced contractile force of the models. This validated new human muscle MPS will benefit development of drugs against muscle wasting diseases. Moreover, our Matrigel 3D bioprinting platform will allow engineering of non-self-organizing complex human 3D MPS.

Highlights

A key to enhance the low translatability of preclinical drug discovery are in vitro human threedimensional (3D) microphysiological systems (MPS)
Musculoskeletal conditions are on a steep rise and are projected to become the most abundant diseases, because prevalent maladies such as cancer and cardiovascular disorders will regress due to healthy life-style, disease prevention efforts and continuously emerging new improved medication[2]
To establish microvalve-based DOD 3D bioprinting of cells suspended in liquid pure Matrigel, a suitable cooling system was developed for the printheads and cartridges of a 3D bioprinter (Fig. 1a), since Matrigel solutions are liquid at low temperature (

Summary

Introduction

A key to enhance the low translatability of preclinical drug discovery are in vitro human threedimensional (3D) microphysiological systems (MPS). Muscle fiber development can be reproduced in vitro using the precursor cells isolated from skeletal muscle biopsies Such 2D tissue culture assays have been used to identify drug candidates regulating growth and maintenance of skeletal muscle such as myostatin pathway blockers[10]. 3D in vitro tissue-like models of contracting skeletal muscle fibers were developed for the screening of compounds regulating contractility[11,12] These human tissue models mimicked pharmacological responses of drugs used in the clinic[13]. Despite its tumor origin, which prevents its clinical application, it is still used for many in vitro tissue engineered models because of missing alternatives providing similar biological cues It became a frequently used key component of the emerging organoid technology, which generates complex human tissues in a dish from stem cells[19]. Despite these successes in 3D tissue culture, Matrigel was rarely used as a bioink and mostly combined with other materials to improve its printability with extrusion bioprinters[20,21,22,23]

Methods

Results

Conclusion