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Three‐dimensional spherical gelatin bubble‐based scaffold improves the myotube formation of H9c2 myoblasts
Author(s) -
Mei Chieh,
Chao ChihWei,
Lin CheWei,
Li Shing Tak,
Wu KuanHan,
Yang KaiChiang,
Yu Jiashing
Publication year - 2019
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26917
Subject(s) - myocyte , myogenesis , microbiology and biotechnology , cell fusion , c2c12 , gelatin , chemistry , cellular differentiation , myosin , biophysics , actin , cell , cytoplasm , cell culture , biology , biochemistry , genetics , gene
Abstract Microenvironmental factors including physical and chemical cues can regulate stem cells as well as terminally differentiated cells to modulate their biological function and differentiation. However, one of the physical cues, the substrate's dimensionality, has not been studied extensively. In this study, the ow‐focusing method with a microuidic device was used to generate gelatin bubbles to fabricate highly ordered three‐dimensional (3D) scaffolds. Rat H9c2 myoblasts were seeded into the 3D gelatin bubble‐based scaffolds and compared to those grown on 2D gelatin‐coating substrates to demonstrate the influences of spatial cues on cell behaviors. Relative to cells on the 2D substrates, the H9c2 myoblasts were featured by a good survival and normal mitochondrial activity but slower cell proliferation within the 3D scaffolds. The cortical actin filaments of H9c2 cells were localized close to the cell membrane when cultured on the 2D substrates, while the F‐actins distributed uniformly and occupied most of the cell cytoplasm within the 3D scaffolds. H9c2 myoblasts fused as multinuclear myotubes within the 3D scaffolds without any induction but cells cultured on the 2D substrates had a relatively lower fusion index even differentiation medium was provided. Although there was no difference in actin α 1 and myosin heavy chain 1, H9c2 cells had a higher myogenin messenger RNA level in the 3D scaffolds than those of on the 2D substrates. This study reveals that the dimensionality influences differentiation and fusion of myoblasts.