ISSN 1662-4009 (online)

ESPE Yearbook of Paediatric Endocrinology (2020) 17 5.15 | DOI: 10.1530/ey.17.5.15

ESPEYB17 5. Bone, Growth Plate and Mineral Metabolism Advances in Skeletal Biology (4 abstracts)

5.15. A simple method based on confocal microscopy and thick sections recognizes seven subphases in growth plate chondrocytes

Fernández-Iglesias Á , Fuente R , Gil-Peña H , Alonso-Duran L , García-Bengoa M & Santos F et al.


Division of Pediatrics, Department of Medicine, Faculty of Medicine, University of Oviedo, Oviedo, Asturias, Spain


To read the full abstract: Sci Rep 2020;10:6935.

In brief: A well-defined and optimized protocol using confocal microscopy allowed novel insight into growth plate chondrocyte differentiation by quantitative analysis of cellular characteristics of murine growth plate samples.

Commentary: The growth plate is a complex tissue with a unique morphology characterized by its anisotropic cellular organization. Limited sample availability, challenging isolation procedures and complex methodologies contributed to the scarce knowledge on growth plate chondrocyte differentiation, which is limited to single studies in recent years. Here, Fernández-Iglesia et al. developed a methodological approach for the standardized characterization of cell volume, cell shape and cytoplasm density during chondrocyte maturation by stringent optimization of protocols for confocal microscopy of growth plate tissue. The authors applied their novel protocol to identify seven clusters of growth plate chondrocytes in murine samples with specific cellular characteristics regarding cell volume, shape and cytoplasmic density. Interestingly, distinct clusters were observed in the proliferative zone, contradicting previous assumptions on homogenous cellular characteristics of proliferative chondrocytes. While this finding underlines the complexity of cellular subpopulations in growth plate cartilage, this work is of special value for providing detailed and comprehensive methodologies including a comparison of fixation techniques to preserve hypertrophic chondrocytes. Further, emphasis was given on reproducibility, feasibility and standardizations, such as by scoring systems for sample quality scoring. Thus, the availability of standardized growth plate imaging methodologies, such as the suggested protocol, might contribute to more detailed investigations of the cellular basis of linear growth, especially in disease models with focus on growth plate alterations and short stature.

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