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Differentiation of Human Induced Pluripotent Stem Cells to Chondrocytes

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1340)

Abstract

Human induced pluripotent stem (iPS) cells are relevant tools for modeling human skeletal development and disease, and represent a promising source of patient-specific cells for the regeneration of skeletal tissue, such as articular cartilage. Devising efficient and reproducible strategies, which closely mimic the physiological chondrogenic differentiation process, will be necessary to generate functional chondrocytes from human iPS cells. Our previous study demonstrated the generation of chondrogenically committed human iPS cells via the enrichment of a mesenchymal-like progenitor population, application of appropriate high-density culture conditions, and stimulation with bone morphogenetic protein-2 (Bmp-2). The differentiated iPS cells showed temporal expression of cartilage genes and the accumulation of a cartilaginous extracellular matrix in vitro. In this chapter, we provide detailed methodologies for the differentiation of human iPS cells to the chondrogenic lineage and describe protocols for the analysis of chondrogenic differentiation.

Key words

Human induced pluripotent stem cells Mesenchymal-like progenitor stem cells Chondrogenic differentiation Micromass culture Bmp-2 
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Notes

Acknowledgements

RMG acknowledges the support of the State of Connecticut Stem Cell Program/Department of Public Health (grants nos. 10SCA36, 13-SCA-UCHC-11). HD acknowledges the support of the State of Connecticut Stem Cell Program/Department of Public Health (Grant #11SCB08). The University of Connecticut Stem Cell Core Facility and the Flow Cytometry Center provided technical support.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Orthopaedic Surgery, MC4037UConn HealthFarmingtonUSA
  2. 2.Stem Cell InstituteUConn HealthFarmingtonUSA
  3. 3.Department of Genetics and Genome SciencesUConn HealthFarmingtonUSA

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