Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels
Autor: | Gio C. Suh, Joon Young Kim, Megan L. McCain, Holly A. Huber, Alyssa A. Viscio, Julie B. Strickland, Clara Hua, Nicholas A. Geisse, Archana Bettadapur, Evelyn R. Wang |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Time Factors food.ingredient Myoblasts Skeletal Muscle Fibers Skeletal Cell Culture Techniques macromolecular substances 02 engineering and technology Microscopy Atomic Force Muscle Development Gelatin Article Cell Line Mice 03 medical and health sciences food medicine Animals Myocyte Dimethylpolysiloxanes Muscle Skeletal Multidisciplinary Tissue Engineering biology Chemistry Myogenesis technology industry and agriculture Skeletal muscle Cell Differentiation Hydrogels Anatomy 021001 nanoscience & nanotechnology Fibronectins Cell biology Fibronectin 030104 developmental biology medicine.anatomical_structure Cell culture Self-healing hydrogels biology.protein 0210 nano-technology C2C12 |
Zdroj: | Scientific Reports |
ISSN: | 2045-2322 |
Popis: | 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. |
Databáze: | OpenAIRE |
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