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Organotypic culture, a powerful model for studying rat and mouse fetal testis development

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Abstract

The key role of the fetal testis in the masculinization of genital organs has been known for a long time. More recently, the observed increases in male reproductive disorders has been postulated to be the result of changes in fetal and neonatal testis development in response to increasing environmental pollution. However, few tools are available for studying fetal testis development and the effects of physiological or toxic substances. We have developed an organ culture system in which rat fetal testis is grown on a filter floating on a synthetic medium containing no serum, hormones or biological factors. In this study, we have compared the long-term morpho-functional development of the various testicular cell types in this system with that observed in vivo and have extended this system to the mouse. Rat Leydig, Sertoli and germ cells and macrophages develop normally over a period of 1–2 weeks in this system. Fewer cells are produced than in vivo but the level of differentiated function is similar. Germ cells, which are difficult to culture in vitro, resume mitosis after a quiescent period, at the same time as in vivo. Similar results have been obtained with mouse fetuses, except that Leydig cells dedifferentiate in vitro if the testis is explanted after 13.5 days post conception. Testicular architecture and intercellular communication are sufficiently preserved for the development of the main fetal and neonatal testicular cell types in vitro with no added factors. Our floating-filter organotypic culture system in synthetic medium therefore allows the morpho-functional development of somatic and germ cells in fetal testis explants taken at all developmental stages in rat and at early stages in mouse. This method is potentially useful for studies of the effects of various factors, and of xenobiotics, in particular.

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Acknowledgements

We are grateful to C. Levacher, C. Racine and E. Moreau for profitable discussions, to P. Flament for animal care, to C. Duquenne for expert technical assistance with the testosterone RIA and to A. Gouret for skillful secretarial assistance. We also thank G. Defaye, A. Payne and the late J.M. Saez for providing antibodies and F. Guillou for transferrin RIA.

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Authors and Affiliations

  1. Laboratory of Differentiation and Radiobiology of the Gonads, Research Unit on Gametogenesis and Genotoxicity, Université Paris 7; CEA; INSERM U566, DSV/DRR/SEGG/LDRG, BP6 - 92265, Fontenay aux Roses, France

    Gabriel Livera, Géraldine Delbes, Catherine Pairault, Virginie Rouiller-Fabre & René Habert

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  1. Gabriel Livera
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  2. Géraldine Delbes
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  3. Catherine Pairault
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  4. Virginie Rouiller-Fabre
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  5. René Habert
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Correspondence to Gabriel Livera.

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This work was supported by the Université Paris 7, Commissariat à l’Energie Atomique (CEA) and the Institut National de la Santé et de la Recherche Médicale (INSERM) and by grant Agence Francaise de Sécurité Sanitaire et Environnementale (AFSSE). G. Delbes is the recipient of a fellowship from the Ministère de la Recherche et de la Technologie and from the Association pour la Recherche contre le Cancer.

This work is dedicated to the memory of José Maria Saez in recognition of his helpful and constant encouragement and discussions over the last decade.

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Livera, G., Delbes, G., Pairault, C. et al. Organotypic culture, a powerful model for studying rat and mouse fetal testis development. Cell Tissue Res 324, 507–521 (2006). https://doi.org/10.1007/s00441-006-0167-7

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