Uniaxial Stretching of Cell-Laden Microfibers for Promoting C2C12 Myoblasts Alignment and Myofibers Formation

ACS Appl Mater Interfaces. 2020 Jan 15;12(2):2162-2170. doi: 10.1021/acsami.9b22103. Epub 2020 Jan 2.

Abstract

Fiber-shaped cellular constructs have attracted increasing attention in the regeneration of blood vessels, nerve networks, and skeletal myofibers. Nevertheless, the generation of functional fiber-shaped cellular constructs suffers from limited appropriate microfiber-based fabrication approaches and the maintenance of regenerated tissue functions. Herein, we demonstrate a silicone-tube-based coagulant bath free method to fabricate tens of centimeters long cell-laden microfibers using single UV exposure without pretreatment of nozzles or microchannels. By modulating the exposure time, the gelatin methacrylate microfibers with tissue-like microstructures and mechanical properties are obtained. Then, a culture system integrated with a pillar well-array based stretching device is used to apply uniaxial stretching with various strain ratios in situ to cell-laden microfibers in a 60 mm petri dish. Cells with improved spreading, elongation, and alignment are obtained under uniaxial stretching. Moreover, the promotional effects of uniaxial stretching on the differentiation of C2C12 myoblasts, the formation, and contractility of myofibers become more pronounced with increasing strain ratio and achieve saturation level as strain ratio up to ∼35%.

Keywords: cellular alignment; gelatin methacrylate (GelMA); hydrogel microfibers; skeletal muscle regeneration; uniaxial stretching.

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Line
  • Cell Survival
  • Fluorescence
  • Mice
  • Muscle Fibers, Skeletal / cytology*
  • Myoblasts / cytology*
  • Sarcomeres / metabolism
  • Stress, Mechanical*
  • Swine