title: Recapitulation of physiological spatiotemporal signals promotes in vitro formation of phenotypically stable human articular cartilage.
abstract: Standard isotropic culture fails to recapitulate the spatiotemporal gradients present during native development. Cartilage grown from human mesenchymal stem cells (hMSCs) is poorly organized and unstable in vivo. We report that human cartilage with physiologic organization and in vivo stability can be grown in vitro from self-assembling hMSCs by implementing spatiotemporal regulation during induction. Self-assembling hMSCs formed cartilage discs in Transwell inserts following isotropic chondrogenic induction with transforming growth factor β to set up a dual-compartment culture. Following a switch in the basal compartment to a hypertrophic regimen with thyroxine, the cartilage discs underwent progressive deep-zone hypertrophy and mineralization. Concurrent chondrogenic induction in the apical compartment enabled the maintenance of functional and hyaline cartilage. Cartilage homeostasis, chondrocyte maturation, and terminal differentiation markers were all up-regulated versus isotropic control groups. We assessed the in vivo stability of the cartilage formed under different induction regimens. Cartilage formed under spatiotemporal regulation in vitro resisted endochondral ossification, retained the expression of cartilage markers, and remained organized following s.c. implantation in immunocompromised mice. In contrast, the isotropic control groups underwent endochondral ossification. Cartilage formed from hMSCs remained stable and organized in vivo. Spatiotemporal regulation during induction in vitro recapitulated some aspects of native cartilage development, and potentiated the maturation of self-assembling hMSCs into stable and organized cartilage resembling the native articular cartilage.
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Johnathan J
authors.identifier: 0000-0002-5170-154X
authors.initials: JJ
authors.lastname: Ng
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Yiyong
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Bernhard
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Samuel
authors.initials: S
authors.lastname: Robinson
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: Aonnicha
authors.initials: A
authors.lastname: Burapachaisri
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032.
authors.forename: X Edward
authors.initials: XE
authors.lastname: Guo
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY 10032; gv2131@columbia.edu.|Department of Medicine, Columbia University, New York, NY 10032.
authors.forename: Gordana
authors.identifier: 0000-0002-9382-1574
authors.initials: G
authors.lastname: Vunjak-Novakovic
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002358:Cartilage, Articular; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology* / Q000276:immunology / Q000637:transplantation; D020219:Chondrogenesis / Q000187:drug effects* / Q000502:physiology; D024042:Collagen Type I / Q000235:genetics / Q000276:immunology; D015194:Diffusion Chambers, Culture; D005260:Female; D015870:Gene Expression; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D016513:Mice, SCID; D010012:Osteogenesis / Q000187:drug effects* / Q000502:physiology; D061251:Primary Cell Culture; D013974:Thyroxine / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014183:Transplantation, Heterologous
chemical_list: D015415:Biomarkers; D024042:Collagen Type I; D016212:Transforming Growth Factor beta; D013974:Thyroxine
keywords: biomimetic; cartilage development; cartilage repair; regenerative medicine; tissue engineering