Co-culture of dedifferentiated and primary human chondrocytes obtained from cadaveric donor enhance the histological qua
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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 Anell Olivos-Meza . Cristina Velasquillo Martı´nez . Brenda Olivos Dı´az . Carlos Landa-Solı´s . Mats Brittberg . Raul Pichardo Bahena . Carmina Ortega Sanchez . Valentin Martı´nez . Enrique Alvarez Lara . Jose´ Clemente Ibarra-Ponce de Leo´n Received: 21 December 2016 / Accepted: 26 May 2017 Ó Springer Science+Business Media Dordrecht 2017
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-GideÒ), and pellet of non-cocultured (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-cocultured group was 4.37 (SD ±4.71), while cocultured 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
A. Olivos-Meza Orthopedic Sports Medicine and Arthroscopy Service, Instituto Nacional de Rehabilitacio´n, Mexico City, Mexico
M. Brittberg Region Halland Orthopaedics, Kungsbacka Hospital, Kungsbacka, Sweden
C. Landa-Solı´s C. Ortega Sanchez V. Martı´nez Tissue Engineering, Cell Therapy and Regenerative Medicine Unit, Instituto Nacional de Rehabilitacio´n, Mexico City, Mexico
R. Pichardo Bahena E. Alvarez Lara Pathology Service, Instituto Nacional de Rehabilitacio´n, Mexico City, Mexico
B. Olivos Dı´az Universidad Nacional Auto´noma de Me´xico, Mexico City, Mexico
J. C. Ibarra-Ponce de Leo´n (&) Instituto Nacional de Rehabilitacio´n, Av Me´xicoXochimilco No. 289, Arenal de Guadalupe, ZC, 14389 Mexico City, Mexico e-mail: [email protected]
B. Olivos Dı´az Instituto Nacional de Rehabilitacio´n, Mexico City, Mexico
C. Velasquillo Martı´nez Subdirection of technological research, Instituto Nacional de Rehabilitacio´n, Mexico City, Mexico
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