Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

Citations Over TimeTop 10% of 2016 papers

Abstract

In vitro models of skeletal muscle are critically needed to elucidate disease mechanisms, identify therapeutic targets, and test drugs pre-clinically. However, culturing skeletal muscle has been challenging due to myotube delamination from synthetic culture substrates approximately one week after initiating differentiation from myoblasts. In this study, we successfully maintained aligned skeletal myotubes differentiated from C2C12 mouse skeletal myoblasts for three weeks by utilizing micromolded (μmolded) gelatin hydrogels as culture substrates, which we thoroughly characterized using atomic force microscopy (AFM). Compared to polydimethylsiloxane (PDMS) microcontact printed (μprinted) with fibronectin (FN), cell adhesion on gelatin hydrogel constructs was significantly higher one week and three weeks after initiating differentiation. Delamination from FN-μprinted PDMS precluded robust detection of myotubes. Compared to a softer blend of PDMS μprinted with FN, myogenic index, myotube width, and myotube length on μmolded gelatin hydrogels was similar one week after initiating differentiation. However, three weeks after initiating differentiation, these parameters were significantly higher on μmolded gelatin hydrogels compared to FN-μprinted soft PDMS constructs. Similar results were observed on isotropic versions of each substrate, suggesting that these findings are independent of substrate patterning. Our platform enables novel studies into skeletal muscle development and disease and chronic drug testing in vitro.

Related Papers

• → Photolithographic Patterning of C2C12 Myotubes using Vitronectin as Growth Substrate in Serum‐Free Medium(2007)69 cited
• → Graphene‐Based Patterning and Differentiation of C2C12 Myoblasts(2013)53 cited
• → Replication of prions in differentiated muscle cells(2014)6 cited
• → Overexpression of Six1 leads to retardation of myogenic differentiation in C2C12 myoblasts(2012)4 cited
• → Effects of Chaenomelis Fructus Extract on the regulation of myoblasts differentiation and the expression of biogenetic factors in C2C12 myotubes(2019)5 cited