Protocol for the fabrication of enzymatically crosslinked gelatin microchannels for microfluidic cell culture.
We have developed a technique for fabricating microfluidic devices from gelatin using a natural crosslinking process. By producing reusable poly(dimethyl siloxane) molds using standard photolithography, gelatin can be molded into microchannel geometries. The gelatin is crosslinked with the naturally occurring enzyme transglutaminase via a straightforward process that can produce devices suitable for cell culture. The protocol takes approximately 1 day from the start of gelatin preparation to cell seeding. Using these devices, the effects of both the extracellular matrix and soluble factors on cellular behavior and differentiation can be studied in microenvironments that more closely mimic the in vivo environment.
Paguirigan
Amy L
AL
Department of Biomedical Engineering, University of Wisconsin, Engineering Centers Building, 1150 Engineering Dr., Madison, Wisconsin 53706, USA.
Beebe
David J
DJ
D018929:Cell Culture Techniques / Q000379:methods*; D002468:Cell Physiological Phenomena; D003432:Cross-Linking Reagents; D004798:Enzymes; D005109:Extracellular Matrix / Q000502:physiology; D005780:Gelatin / Q000737:chemistry* / Q000235:genetics; D005782:Gels; D015870:Gene Expression; D005973:Glutamine / Q000032:analysis; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D044085:Microfluidics
D003432:Cross-Linking Reagents; D004798:Enzymes; D005782:Gels; D005973:Glutamine; D005780:Gelatin