An in vitro prototype of a porcine biomimetic testis-like cell culture system： a novel tool for the study of reassembled Sertoli and Leydig cells  

作　　者：Iva Arato Giovanni Luca Francesca Mancuso Catia Bellucci Cinzia Lilli Mario Calvitti Barbara C Hansen Domenico Milardi Giuseppe Grande Riccardo Calafiore 

机构地区：[1]Department of Experimental Medicine, University of Perugia, Perugia 06100, Italy [2]Division of Medical Andrology and Endocrinology of Reproduction, University of Perugia and Saint Mary Hospital, Terni 05100, Italy [3]Department of Internal Medicine and Pediatrics, Morsani College of Medicine, University of South Florida, Tampa 33612, FL, USA [4]Division of Endocrinology, Fondazione Policlinico Universitario 'A. Gemelli', 00168 Rome, Italy [5]Department of Medicine, University of Perugia, Perugia 06100, Italy

出　　处：《Asian Journal of Andrology》2018年第2期160-165,共6页亚洲男性学杂志（英文版）

摘　　要：At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors （testicular cross-talk）, mainly orchestrated by gonadotropins such as luteinizing hormone （LH） and follicle-stimulating hormone （FSH） that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay （ELISA） for anti-Mullerian hormone （AMH）, inhibin B, and testosterone secretion in the medium, and Real-lime PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3βHSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors （testicular cross-talk）, mainly orchestrated by gonadotropins such as luteinizing hormone （LH） and follicle-stimulating hormone （FSH） that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay （ELISA） for anti-Mullerian hormone （AMH）, inhibin B, and testosterone secretion in the medium, and Real-lime PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3βHSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.

关 键 词：human gonadotropins Leydig cells prepubertal biomimetic testis Sertoli cells 

分 类 号：Q579.11[生物学—生物化学] G12[文化科学]