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Morphological, genetic and phenotypic comparison between human articular chondrocytes and cultured chondrocytes

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Abstract

Articular cartilage is an avascular and aneural tissue with limited capacity for regeneration. On large articular lesions, it is recommended to use regenerative medicine strategies, like autologous chondrocyte implantation. There is a concern about morphological changes that chondrocytes suffer once they have been isolated and cultured. Due to the fact that there is little evidence that compares articular cartilage chondrocytes with cultured chondrocytes, in this research we proposed to obtain chondrocytes from human articular cartilage, compare them with themselves once they have been cultured and characterize them through genetic, phenotypic and morphological analysis. Knee articular cartilage samples of 10 mm were obtained, and each sample was divided into two fragments; a portion was used to determine gene expression, and from the other portion, chondrocytes were obtained by enzymatic disaggregation, in order to be cultured and expanded in vitro. Subsequently, morphological, genetic and phenotypic characteristics were compared between in situ (articular cartilage) and cultured chondrocytes. Obtained cultured chondrocytes were rounded in shape, possessing a large nucleus with condensed chromatin and a clear cytoplasm; histological appearance was quite similar to typical chondrocyte. The expression levels of COL2A1 and COL10A1 genes were higher in cultured chondrocytes than in situ chondrocytes; moreover, the expression of COL1A1 was almost undetectable on cultured chondrocytes; likewise, COL2 and SOX9 proteins were detected by immunofluorescence. We concluded that chondrocytes derived from adult human cartilage cultured for 21 days do not tend to dedifferentiate, maintaining their capacity to produce matrix and also retaining their synthesis capacity and morphology.

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Acknowledgments

The A022-2014 budgetary program supported this work. We appreciate the collaboration of Ricardo Martínez Flores, student of medicine at Escuela Medico Militar.

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Authors and Affiliations

  1. Laboratorio de Biología Celular y Tisular, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, 11200, Ciudad de México, Mexico

    Mónica Maribel Mata-Miranda & Claudia María Martinez-Martinez

  2. CIBA-Tlaxcala, Instituto Politécnico Nacional, 90700, Tepetitla, Tlaxcala, Mexico

    Mónica Maribel Mata-Miranda & Gustavo Jesús Vázquez-Zapién

  3. Laboratorio de Embriología, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, Cerrada de Palomas S/N, Lomas de Sotelo, Miguel Hidalgo, 11200, Ciudad de México, Mexico

    Jesús Emmanuel Noriega-Gonzalez & Gustavo Jesús Vázquez-Zapién

  4. Laboratorio Multidisciplinario de Investigación, Escuela Militar de Graduados de Sanidad, Universidad del Ejército y Fuerza Aérea, 11200, Ciudad de México, Mexico

    Luis Enrique Paredes-Gonzalez

Authors
  1. Mónica Maribel Mata-Miranda
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  2. Claudia María Martinez-Martinez
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  3. Jesús Emmanuel Noriega-Gonzalez
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  4. Luis Enrique Paredes-Gonzalez
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  5. Gustavo Jesús Vázquez-Zapién
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Corresponding author

Correspondence to Gustavo Jesús Vázquez-Zapién.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Mata-Miranda, M.M., Martinez-Martinez, C.M., Noriega-Gonzalez, J.E. et al. Morphological, genetic and phenotypic comparison between human articular chondrocytes and cultured chondrocytes. Histochem Cell Biol 146, 183–189 (2016). https://doi.org/10.1007/s00418-016-1437-4

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  • DOI: https://doi.org/10.1007/s00418-016-1437-4

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