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Co-culture of dedifferentiated and primary human chondrocytes obtained from cadaveric donor enhance the histological quality of repair tissue: an in-vivo animal study

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Abstract

To compare the quality of the repair tissue in three-dimensional co-culture of human chondrocytes implanted in an in vivo model. Six cadaveric and five live human donors were included. Osteochondral biopsies from the donor knees were harvested for chondrocyte isolation. Fifty percent of cadaveric chondrocytes were expanded until passage-2 (P2) while the remaining cells were cryopreserved in passage-0 (P0). Fresh primary chondrocytes (P0f) obtained from live human donors were co-cultured. Three-dimensional constructs were prepared with a monolayer of passage-2 chondrocytes, collagen membrane (Geistlich Bio-Gide®), and pellet of non-co-cultured (P2) or co-cultured chondrocytes (P2 + P0c, P2 + P0f). Constructs were implanted in the subcutaneous tissue of athymic mice and left for 3 months growth. Safranin-O and Alcian blue staining were used to glycosaminoglycan content assessment. Aggrecan and type-II collagen were evaluated by immunohistochemistry. New-formed tissue quality was evaluated with an adaptation of the modified O’Driscoll score. Histological quality of non-co-cultured group was 4.37 (SD ±4.71), while co-cultured groups had a mean score of 8.71 (SD ±3.98) for the fresh primary chondrocytes and 9.57 (SD ±1.27) in the cryopreserved chondrocytes. In immunohistochemistry, Co-culture groups were strongly stained for type-II and aggrecan not seen in the non-co-cultured group. It is possible to isolate viable chondrocytes from cadaveric human donors in samples processed in the first 48-h of dead. There is non-significant difference between the numbers of chondrocytes isolated from live or cadaveric donors. Cryopreservation of cadaveric primary chondrocytes does not alter the capability to form cartilage like tissue. Co-culture of primary and passaged chondrocytes enhances the histological quality of new-formed tissue compared to non-co-cultured cells.

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Acknowledgements

This work is supported by the Mexican Council of Science (CONACyT) Grant SALUD-PDCPN-2013-01-215138; Technology, Science and Innovation Secretary Grants: SECITI 079 BIS/2013 and SECITI/INR/GOB-25/2013. The authors wish specialty to thank to Biograft of México Musculoskeletal Tissue and Skin Bank; Blood Bank & Pathology Service at the National Institute of Rehabilitation; and Autonomous National University of Mexico (UNAM). We also appreciate the support and collaboration of Veterinary Facility at the National Institute of Rehabilitation: Hugo-Lecona DMV and Javier-Perez DMV.

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Authors and Affiliations

  1. Orthopedic Sports Medicine and Arthroscopy Service, Instituto Nacional de Rehabilitación, Mexico City, Mexico

    Anell Olivos-Meza

  2. Tissue Engineering, Cell Therapy and Regenerative Medicine Unit, Instituto Nacional de Rehabilitación, Mexico City, Mexico

    Carlos Landa-Solís, Carmina Ortega Sanchez & Valentin Martínez

  3. Universidad Nacional Autónoma de México, Mexico City, Mexico

    Brenda Olivos Díaz

  4. Instituto Nacional de Rehabilitación, Mexico City, Mexico

    Brenda Olivos Díaz

  5. Region Halland Orthopaedics, Kungsbacka Hospital, Kungsbacka, Sweden

    Mats Brittberg

  6. Pathology Service, Instituto Nacional de Rehabilitación, Mexico City, Mexico

    Raul Pichardo Bahena & Enrique Alvarez Lara

  7. Instituto Nacional de Rehabilitación, Av México-Xochimilco No. 289, Arenal de Guadalupe, ZC, 14389, Mexico City, Mexico

    José Clemente Ibarra-Ponce de León

  8. Subdirection of technological research, Instituto Nacional de Rehabilitación, Mexico City, Mexico

    Cristina Velasquillo Martínez

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  1. Anell Olivos-Meza
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  2. Cristina Velasquillo Martínez
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  3. Brenda Olivos Díaz
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  4. Carlos Landa-Solís
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  5. Mats Brittberg
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  6. Raul Pichardo Bahena
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  7. Carmina Ortega Sanchez
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  8. Valentin Martínez
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  9. Enrique Alvarez Lara
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  10. José Clemente Ibarra-Ponce de León
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Correspondence to José Clemente Ibarra-Ponce de León.

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Olivos-Meza, A., Velasquillo Martínez, C., Olivos Díaz, B. et al. Co-culture of dedifferentiated and primary human chondrocytes obtained from cadaveric donor enhance the histological quality of repair tissue: an in-vivo animal study. Cell Tissue Bank 18, 369–381 (2017). https://doi.org/10.1007/s10561-017-9635-4

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  • DOI: https://doi.org/10.1007/s10561-017-9635-4

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