Encapsulation of Human Articular Chondrocytes into 3D Hydrogel: Phenotype and Genotype Characterization

  • Rui C. PereiraEmail author
  • Chiara Gentili
  • Ranieri Cancedda
  • Helena S. Azevedo
  • Rui L. Reis
Part of the Methods in Molecular Biology book series (MIMB, volume 695)


This chapter is intended to provide a summary of the current materials used in cell encapsulation technology as well as methods for evaluating the performance of cells encapsulated in a polymeric matrix. In particular, it describes the experimental procedure to prepare a hydrogel matrix based on natural polymers for encapsulating and culturing human articular chondrocytes with the interest in cartilage regeneration. Protocols to evaluate the viability, proliferation, differentiation, and matrix production of embedded cells are also described and include standard protocols such as the MTT and [3H] Thymidine assays, reverse transcription polymerase chain reaction (RT-PCR) technique, histology, and immunohistochemistry analysis. The assessment of cell distribution within the 3D hydrogel construct is also described using APoTome analysis.

Key words

Natural polymers Hydrogels Gelation Chondrocyte encapsulation Cell culture and survival within 3D matrix Cell-based technology Cartilage regeneration 



The authors would like to thank Recco Orthopedic staff members for helpful discussions, particularly to Dr. Michele Grandizio; patients for biopsy material donation, as well as to Mrs Daniela Marubbi and Monica Scaranari for technical assistance with histological sample processing. This work was supported by funds from the Italian MUR (FIRB-Tissuenet project), the European Union funded STREP Project HIPPOCRATES (NMP3-CT-2003-505758), and the European NoE EXPERTISSUES (NMP3-CT-2004-500283).


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Rui C. Pereira
    • 1
    • 2
    • 3
    Email author
  • Chiara Gentili
    • 3
  • Ranieri Cancedda
    • 3
  • Helena S. Azevedo
    • 1
    • 2
  • Rui L. Reis
    • 1
    • 2
  1. 1.Biomaterials, Biodegradables and BiomimeticsUniversity of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineGuimarãesPortugal
  2. 2.PTGovernment Associated LaboratoryInstitute for Biotechnology and BioengineeringGuimarãesPortugal
  3. 3.Dipartimento di Biologia, Oncologia e GeneticaUniversita di Genoval & Istituto Nazionale per la Ricerca sul CancroGenovaItaly

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