title: Tympanic membrane organ culture using cell culture well inserts engrafted with tympanic membrane tissue explants.
abstract: Tissue engineering approaches using growth factors and various materials for repairing chronic perforations of the tympanic membrane are being developed, but there are surprisingly few relevant tissue culture models available to test new treatments. Here, we present a simple three-dimensional model system based on micro-dissecting the rat tympanic membrane umbo and grafting it into the membrane of a cell culture well insert. Cell outgrowth from the graft produced sufficient cells to populate a membrane of similar surface area to the human tympanic membrane within 2 weeks. Tissue grafts from the annulus region also showed cell outgrowth but were not as productive. The umbo organoid supported substantial cell proliferation and migration under the influence of keratinocyte growth medium. Cells from umbo grafts were enzymatically harvested from the polyethylene terephthalate (PET) membrane for expansion in routine culture and cells could be harvested consecutively from the same graft over multiple cycles. We used harvested cells to test cell migration properties and to engraft a porous silk scaffold material as proof-of-principle for tissue engineering applications. This model is simple enough to be widely adopted for tympanic membrane regeneration studies and has promise as a tissue-equivalent model alternative to animal testing.
authors.affiliation: Ear Sciences Centre, School of Surgery, University of Western Australia, Perth, Australia.|Ear Science Institute Australia, Perth, Australia.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Liew
authors.affiliation: Division of Medicine, University College London, UK.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Day
authors.affiliation: Ear Sciences Centre, School of Surgery, University of Western Australia, Perth, Australia.|Ear Science Institute Australia, Perth, Australia.|Centre for Cell Therapy and Regenerative Medicine, University of Western Australia, Perth, Australia.
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Dilley
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement; D002478:Cells, Cultured; D006801:Humans; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014432:Tympanic Membrane / Q000166:cytology*
keywords: explant culture; silk fibroin; tympanic membrane