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J Orthop Res. 2012 Nov;30(11):1753-9. doi: 10.1002/jor.22123. Epub 2012 Apr 19.

Sequential differentiation of mesenchymal stem cells in an agarose scaffold promotes a physis-like zonal alignment of chondrocytes.

Author information

  • 1Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 10, Kent Ridge Crescent, Singapore 119260, Singapore.

Erratum in

  • J Orthop Res. 2013 Oct;31(10):1676. Hua, See Kwee [corrected to See, Kwee Hua]; Zheng, Yang [corrected to Yang, Zheng]; Po, James Hui Hoi [corrected to Hui, James Hoi Po]; Hin, Lee Eng [corrected to Lee, Eng Hin].


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.

Copyright © 2012 Orthopaedic Research Society.

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