Photolithographic patterning of C2C12 myotubes using vitronectin as growth substrate in serum-free medium.
The C2C12 cell line is frequently used as a model of skeletal muscle differentiation. In our serum-free defined culture system, differentiation of C2C12 cells into myotubes required surface-bound signals such as substrate-adsorbed vitronectin or laminin. On the basis of this substrate requirement of myotube formation, we developed a photolithography-based method to pattern C2C12 myotubes, where myotubes formed exclusively on vitronectin surface patterns. We have determined that the optimal line width to form single myotubes is approximately 30 mum. To illustrate a possible application of this method, we patterned myotubes on the top of commercial substrate-embedded microelectrodes. In contrast to previous experiments where cell patterning was achieved by selective attachment of the cells to patterned surfaces in a medium that contained all of the factors necessary for differentiation, this study illustrates that surface patterning of a signaling molecule, which is essential for skeletal muscle differentiation in a defined system, can result in the formation of aligned myotubes on the patterns. This technique is being developed for applications in cell biology, tissue engineering, and robotics.
Molnar
Peter
P
Nanoscience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, Florida 32826, USA. pmolnar@mail.ucf.edu
Wang
Weishi
W
Natarajan
Anupama
A
Rumsey
John W
JW
Hickman
James J
JJ
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D020099:Coated Materials, Biocompatible / Q000008:administration & dosage / Q000737:chemistry; D016895:Culture Media, Serum-Free; D051379:Mice; D032342:Muscle Cells / Q000166:cytology* / Q000187:drug effects; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000187:drug effects; D010781:Photography / Q000379:methods; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*; D019096:Vitronectin / Q000008:administration & dosage* / Q000737:chemistry
D020099:Coated Materials, Biocompatible; D016895:Culture Media, Serum-Free; D019096:Vitronectin