Integration and application of vitrified collagen in multilayered microfluidic devices for corneal microtissue culture.
This paper describes the fabrication and application of microfluidic devices containing collagen vitrigel (CV) used as both a functional and sacrificial cell growth substrate for the development of corneal microtissue patches. Within the device, vacuum fixation of the CV in a dehydrated state enables quick integration with standard multilayer soft lithographic techniques, while on-chip rehydration results in a gel-like collagen substrate for microfluidic cell culture. Fluidic connectivity to both the apical and basal side of the CV permits bilayered culture of epithelium and supporting stromal cell layers. In addition, microfluidic introduction of a collagenase etching media enables sacrificial degradation of the supporting CV membrane for development of barrier tissue constructs containing minimal synthetic substrate. The utility of this platform was evaluated by miniaturizing the standard transepithelial permeability (TEP) assay in order to measure the integrity of an array of corneal tissue micropatches.
Johns Hopkins University, Department of Biomedical Engineering, 3400 N. Charles St., Clark Hall, Baltimore, MD, USA. cpuleo@jhu.edu
Christopher M
CM
Puleo
Winnette
W
McIntosh Ambrose
Toshiaki
T
Takezawa
Jennifer
J
Elisseeff
Tza-Huei
TH
Wang
D000818:Animals; D003094:Collagen / Q000737:chemistry*; D003315:Cornea / Q000166:cytology*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004847:Epithelial Cells / Q000166:cytology; D005782:Gels; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D018613:Microscopy, Confocal; D009757:Nylons / Q000737:chemistry; D011817:Rabbits; D046509:Tissue Culture Techniques / Q000379:methods*
D004129:Dimethylpolysiloxanes; D005782:Gels; D009757:Nylons; C085707:poly(dimethylsiloxane)-polyamide copolymer; D003094:Collagen