title: Sequential differentiation of mesenchymal stem cells in an agarose scaffold promotes a physis-like zonal alignment of chondrocytes.
abstract: Chondrocytes of the epiphyseal growth plate (physis) differentiate and mature in defined linear zones. The current study examines the differentiation of human bone marrow derived mesenchymal stem cells (hBMSCs) into zonal physeal cartilage. hBMSCs were embedded in an agarose scaffold with only the surface of the scaffold in direct contact with the culture medium. The cells were differentiated using a two-step system involving the sequential addition of TGFβ followed by BMP2. The resultant samples displayed a heterogenic population of physis-like collagen type 2 positive cells including proliferating chondrocytes and mature chondrocytes showing hypertrophy, expression of early bone markers and matrix mineralization. Histological analysis revealed a physis-like linear zonal alignment of chondrocytes in varying stages of differentiation. The less mature chondrocytes were seen at the base of the construct while hypertrophic chondrocytes and matrix mineralization was observed closer to the surface of the construct. The described differentiation protocol using hBMSCs in an agarose scaffold can be used to study the factors and conditions that influence the differentiation, proliferation, maturation, and zonal alignment of physeal chondrocytes.
authors.affiliation: Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 10, Kent Ridge Crescent, Singapore 119260, Singapore.
authors.forename: Jacqueline Frida
authors.initials: JF
authors.lastname: Schmitt
authors.forename: Kwee Hua
authors.initials: KH
authors.lastname: See
authors.forename: See Kwee
authors.initials: SK
authors.lastname: Hua
authors.forename: Zheng
authors.initials: Z
authors.lastname: Yang
authors.forename: Yang
authors.initials: Y
authors.lastname: Zheng
authors.forename: James Hoi Po
authors.initials: JH
authors.lastname: Hui
authors.forename: James Hui Hoi
authors.initials: JH
authors.lastname: Po
authors.forename: Eng Hin
authors.initials: EH
authors.lastname: Lee
authors.forename: Lee Eng
authors.initials: LE
authors.lastname: Hin
mesh_terms: D000293:Adolescent; D055396:Bone Morphogenetic Protein 2; D002356:Cartilage / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D019902:Chondrocytes / Q000166:cytology*; D020219:Chondrogenesis; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D054457:Tissue Scaffolds; D016212:Transforming Growth Factor beta
chemical_list: D055396:Bone Morphogenetic Protein 2; D016212:Transforming Growth Factor beta