title: Double-chamber rotating bioreactor for dynamic perfusion cell seeding of large-segment tracheal allografts: comparison to conventional static methods.
abstract: Tracheal transplantation with a long-segment recellularized tracheal allograft has previously been performed without the need for immunosuppressive therapy. Recipients' mesenchymal stromal cells (MSC) and tracheal epithelial cells (TEC) were harvested, cultured, expanded, and seeded on a donor trachea within a bioreactor. Prior techniques used for cellular seeding have involved only static-seeding methods. Here, we describe a novel bioreactor for recellularization of long-segment tracheae. Tracheae were recellularized with epithelial cells on the luminal surface and bone marrow-derived MSC on the external surface. We used dynamic perfusion seeding for both cell types and demonstrate an increase in both cellular counts and homogeneity scores compared with traditional methods. Despite these improvements, orthotopic transplantation of these scaffolds revealed no labeled cells at postoperative day 3 and lack of re-epithelialization within the first 2 weeks. The animals in this study had postoperative respiratory distress and tracheal collapse that was incompatible with life.
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network , Toronto, Canada .
authors.forename: Siba
authors.initials: S
authors.lastname: Haykal
authors.forename: Michael
authors.initials: M
authors.lastname: Salna
authors.forename: Yingzhe
authors.initials: Y
authors.lastname: Zhou
authors.forename: Paula
authors.initials: P
authors.lastname: Marcus
authors.forename: Mostafa
authors.initials: M
authors.lastname: Fatehi
authors.forename: Geoff
authors.initials: G
authors.lastname: Frost
authors.forename: Tiago
authors.initials: T
authors.lastname: Machuca
authors.forename: Stefan O P
authors.initials: SO
authors.lastname: Hofer
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Waddell
mesh_terms: D064591:Allografts; D000818:Animals; D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods; D016377:Organ Transplantation / Q000379:methods; D013552:Swine; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D014132:Trachea / Q000166:cytology* / Q000637:transplantation