A simple and scalable 3D printing methodology for generating aligned and extended human and murine skeletal muscle tissues

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  • Selgin D Cakal
  • Carmen Radeke
  • Juan F Alcala
  • Ditte G Ellman
  • Sarkhan Butdayev
  • Ditte C Andersen
  • Callø, Kirstine
  • Johan U Lind

Preclinical biomedical and pharmaceutical research on disease causes, drug targets, and side effects increasingly relies on in vitromodels of human tissue. 3D printing offers unique opportunities for generating models of superior physiological accuracy, as well as for automating their fabrication. Towards these goals, we here describe a simple and scalable methodology for generating physiologically relevant models of skeletal muscle. Our approach relies on dual-material micro-extrusion of two types of gelatin hydrogel into patterned soft substrates with locally alternating stiffness. We identify minimally complex patterns capable of guiding the large-scale self-assembly of aligned, extended, and contractile human and murine skeletal myotubes. Interestingly, we find high-resolution patterning is not required, as even patterns with feature sizes of several hundred micrometers is sufficient. Consequently, the procedure is rapid and compatible with any low-cost extrusion-based 3D printer. The generated myotubes easily span several millimeters, and various myotube patterns can be generated in a predictable and reproducible manner. The compliant nature and adjustable thickness of the hydrogel substrates, serves to enable extended culture of contractile myotubes. The method is further readily compatible with standard cell-culturing platforms as well as commercially available electrodes for electrically induced exercise and monitoring of the myotubes.

Original languageEnglish
JournalBiomedical Materials (Bristol)
Volume17
Issue number4
ISSN1748-6041
DOIs
Publication statusPublished - 2022

Bibliographical note

Creative Commons Attribution license.

    Research areas

  • Animals, Humans, Hydrogels, Mice, Muscle Fibers, Skeletal, Muscle, Skeletal, Printing, Three-Dimensional, Tissue Engineering/methods

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