Morphological, genetic and phenotypic comparison between human articular chondrocytes and cultured chondrocytes.
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.
Laboratorio de Biología Celular y Tisular, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, 11200, Ciudad de México, Mexico.|CIBA-Tlaxcala, Instituto Politécnico Nacional, 90700, Tepetitla, Tlaxcala, Mexico.
Mónica Maribel
MM
Mata-Miranda
Laboratorio de Biología Celular y Tisular, Escuela Médico Militar, Universidad del Ejército y Fuerza Aérea, 11200, Ciudad de México, Mexico.
Claudia María
CM
Martinez-Martinez
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
JE
Noriega-Gonzalez
Laboratorio Multidisciplinario de Investigación, Escuela Militar de Graduados de Sanidad, Universidad del Ejército y Fuerza Aérea, 11200, Ciudad de México, Mexico.
Luis Enrique
LE
Paredes-Gonzalez
CIBA-Tlaxcala, Instituto Politécnico Nacional, 90700, Tepetitla, Tlaxcala, Mexico. gus1202@hotmail.com.|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. gus1202@hotmail.com.
Gustavo Jesús
GJ
Vázquez-Zapién
D000328:Adult; D002358:Cartilage, Articular / Q000166:cytology* / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism*; D024043:Collagen Type II / Q000032:analysis / Q000235:genetics; D024161:Collagen Type XI / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D010641:Phenotype; D055758:SOX9 Transcription Factor / Q000032:analysis; D055815:Young Adult
C574182:COL11A1 protein, human; C499177:COL2A1 protein, human; D024043:Collagen Type II; D024161:Collagen Type XI; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human
Articular injuries; Characterized chondrocytes; Cultured chondrocytes