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This version published online on April 18, 2006
Journal of Clinical Endocrinology & Metabolism, doi:10.1210/jc.2005-2113
A more recent version of this article appeared on July 1, 2006
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Submitted on September 23, 2005
Accepted on April 6, 2006

Use of organ culture to study the human fetal testis development: effect of retinoic acid

Romain Lambrot*, Hervé Coffigny, Catherine Pairault, Anne-Claire Donnadieu, René Frydman, René Habert, and Virginie Rouiller-Fabre

CEA, DSV/DRR/SEGG/LDRG, Laboratory of Differentiation and Radiobiology of the Gonads, Unit of Gametogenesis and Genotoxicity (R.L., H.C., C.P., R.H., V.R.F.), F-92265 Fontenay-aux-Roses, France; INSERM, U566 (R.L., H.C., C.P., R.H., V.R.F.), F-92265 Fontenay-aux-Roses, France; Univ Paris 7 Denis Diderot (R.L., H.C., C.P., R.H., V.R.F.), U.F.R. of Biology, F-92265 Fontenay-aux-Roses, France; AP-HP, Groupement hospitalier universitaire Sud, Hôpital A. Béclère, Service de Gynécologie-Obstétrique (A.C.D., R.F.), F-92141 Clamart, France; IFR 13 (R.L., H.C., C.P., A.C.D., R.F., R.H., V.R.F.)

* To whom correspondence should be addressed. E-mail: romain.lambrot{at}cea.fr.

Context: In human, the chronology of the testicular development has been extensively studied but the factors implicated in the onset and the regulation of gametogenesis and steroidogenesis remain hardly known.

Objectives: To identify these factors we have developed an organ culture system for human fetal testes recovered during the first trimester (6-12 weeks) of gestation. We first aimed at investigating the characteristics of this system by comparing the in vivo and in vitro gametogenesis and steroidogenesis. In a second time, we used organ culture to investigate the effect on the human testicular functions of retinoic acid (RA), previously described as a regulator of gonadal development in rodents.

Results: Organ culture proved to be an efficient tool for studying the early development of the testicular functions. Indeed, this system was able to maintain satisfactory development of the germ cells and Leydig cells in the absence of any added factor. For older fetuses, the number of germ cells decreased in culture and the LH was necessary to maintain the steroidogenic activity. The addition of 10-6 M RA decreased the total number of germ cells in the fetal testis at all studied stages. This resulted from an increase in apoptosis, which slightly exceeded the increase of proliferation. However, RA had a stimulatory effect on the steroidogenic function for the youngest fetuses over a short period of time by increasing the expression of P450SCC, P450C17 and StAR.

Conclusions: Thus, RA appears as a potential regulator of both gametogenesis and steroidogenesis in human fetal testis. Our organ culture is an interesting tool for studying the effects of various factors on the development of human fetal testis, in particular the effect of hormone-disrupting chemicals.
Key words: human fetus • testis development • organ culture • retinoic acid




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