Fabrication of ultrathin artificial basement membrane for epithelial cell culture
Résumé
Basement membranes are essential for epithelial and endothelial tissue organization. To mimic them, we developed a fabrication method to produce ultrathin membrane of collagen IV and laminin. A honeycomb microframe of thickness 50 µm and compartment-size 400 µm was firstly patterned in polyethylene glycol diacrylate (PEGDA) by using lithography and vacuum assisted UV curing techniques. Then, a monolayer of gelatin nanofibers was electrospun and crosslinked on the microframe to form a secondary structure, i.e. a fiber mesh with much smaller pore sizes, in each of the honeycomb compartments. Finally, a collagen IV-laminin gel layer was deposited and dehydrated, leading to the formation of an ultrathin membrane over a large area with a nanofiber backbone. Such an artificial basement membrane is mechanically stable and fully bio-compatible. It is also semi-permeable which slowed down considerably the diffusion of large size molecules. More importantly, it could be used to improve the monolayer formation of epithelial cells. Thus, this culture device recapitulates the biological basement membrane, permits the epithelial tissue formation, and allows mimicking a more sophisticated cellular microenvironment.
Domaines
Biotechnologies
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Fabrication of ultrathin artificial basement membrane for epithelial cell culture.pdf (670.04 Ko)
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