Human elastic cartilage engineering from cartilage progenitor cells using rotating wall vessel bioreactor |
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Authors: | Takebe T Kobayashi S Kan H Suzuki H Yabuki Y Mizuno M Adegawa T Yoshioka T Tanaka J Maegawa J Taniguchi H |
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Institution: | a Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku, Yokohama, Kanagawa, Japan b Department of Plastic and Reconstructive Surgery, Kanagawa Children's Medical Center, Minami-ku, Yokohama, Japan c Department of Plastic and Reconstructive Surgery, Yokohama City University Graduate School of Medicine, Yokohama, Kanagawa, Japan d Department of Metallurgy and Ceramics Science, Tokyo Institute of Technology, Tokyo, Japan |
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Abstract: | Transplantation of bioengineered elastic cartilage is considered to be a promising approach for patients with craniofacial defects. We have previously shown that human ear perichondrium harbors a population of cartilage progenitor cells (CPCs). The aim of this study was to examine the use of a rotating wall vessel (RWV) bioreactor for CPCs to engineer 3-D elastic cartilage in vitro. Human CPCs isolated from ear perichondrium were expanded and differentiated into chondrocytes under 2-D culture conditions. Fully differentiated CPCs were seeded into recently developed pC-HAp/ChS (porous material consisted of collagen, hydroxyapatite, and chondroitinsulfate) scaffolds and 3-D cultivated utilizing a RWV bioreactor. 3-D engineered constructs appeared shiny with a yellowish, cartilage-like morphology. The shape of the molded scaffold was maintained after RWV cultivation. Hematoxylin and eosin staining showed engraftment of CPCs inside pC-HAp/ChS. Alcian blue and Elastica Van Gieson staining showed of proteoglycan and elastic fibers, which are unique extracellular matrices of elastic cartilage. Thus, human CPCs formed elastic cartilage-like tissue after 3-D cultivation in a RWV bioreactor. These techniques may assist future efforts to reconstruct complicate structures composed of elastic cartilage in vitro. |
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