Mesoscopic hydrogel molding to control the 3D geometry of bioartificial muscle tissues

Nat Protoc. 2009;4(10):1522-34. doi: 10.1038/nprot.2009.155. Epub 2009 Sep 24.

Abstract

This protocol describes a cell/hydrogel molding method for precise and reproducible biomimetic fabrication of three-dimensional (3D) muscle tissue architectures in vitro. Using a high aspect ratio soft lithography technique, we fabricate polydimethylsiloxane (PDMS) molds containing arrays of mesoscopic posts with defined size, elongation and spacing. On cell/hydrogel molding, these posts serve to enhance the diffusion of nutrients to cells by introducing elliptical pores in the cell-laden hydrogels and to guide local 3D cell alignment by governing the spatial pattern of mechanical tension. Instead of ultraviolet or chemical cross-linking, this method utilizes natural hydrogel polymerization and topographically constrained cell-mediated gel compaction to create the desired 3D tissue structures. We apply this method to fabricate several square centimeter large, few hundred micron-thick bioartificial muscle tissues composed of viable, dense, uniformly aligned and highly differentiated cardiac or skeletal muscle fibers. The protocol takes 4-5 d to fabricate PDMS molds followed by 2 weeks of cell culture.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomimetic Materials*
  • Biomimetics / methods*
  • Hydrogel, Polyethylene Glycol Dimethacrylate*
  • Muscle Fibers, Skeletal
  • Muscles*
  • Tissue Culture Techniques

Substances

  • Hydrogel, Polyethylene Glycol Dimethacrylate