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Two-Dimensional Differential Adherence and Myoblast Alignment Driven Fabrication of Engineered C2C12 Muscle Organoids

Published online by Cambridge University Press:  01 February 2011

Tim M. Patz
Affiliation:
Georgia Institute of Technology, School of Material Science & Engineering, 771 Ferst Drive NW, Atlanta, GA 30332–0245, USA
Anand Doraiswamy
Affiliation:
Georgia Institute of Technology, School of Material Science & Engineering, 771 Ferst Drive NW, Atlanta, GA 30332–0245, USA
Roger Narayan
Affiliation:
Georgia Institute of Technology, School of Material Science & Engineering, 771 Ferst Drive NW, Atlanta, GA 30332–0245, USA
Rohit Modi
Affiliation:
US Naval Research Laboratory, 4555 Overlook Ave. Washington, DC 20375 USA
Douglas Chrisey
Affiliation:
US Naval Research Laboratory, 4555 Overlook Ave. Washington, DC 20375 USA
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Abstract

Differential adherent surfaces were used to induce attachment of C2C12 myoblasts that later would form engineered muscle organoids. Narrow channels, 60–400 μm wide, were micromachined in a 2% agarose gel using an ArF laser, and subsequently filled with extracellular matrix. Upon addition of 1 mL of a 2×104 cells/mL myoblast suspension, cells selectively adhered to the ECM lined channels in a non-confluent manner and we monitored their growth at various time points. The adherent myoblasts were flourescently imaged with a propidium iodide live/dead assay, which revealed that the cells were alive within the channels. After 72 hours growth, the myoblasts grew, proliferated and differentiated into myotubes. Myoblasts residing in the narrow channels (60–150 μm wide) aligned parallel to the channel, while myoblasts adhered to the wide channel (400 μm) were randomly aligned within the channel. The fully-grown 1 cm long organoids maintained an aspect ratio on the order of 100:1. These results provide the foundation for future three-dimensional tissue growth using differential adherent

Type
Research Article
Copyright
Copyright © Materials Research Society 2005

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References

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