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

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

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD 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.

Original languageEnglish
Pages (from-to)160-165
Number of pages6
JournalAsian Journal of Andrology
Volume20
Issue number2
DOIs
Publication statusPublished - Jan 1 2018

Fingerprint

Biomimetics
Leydig Cells
Sertoli Cells
Testis
Swine
Cell Culture Techniques
Gonadotropins
Follicle Stimulating Hormone
Luteinizing Hormone
Testosterone
Hormones
FSH Receptors
Messenger RNA
Aromatase
Organ Culture Techniques
Male Infertility
Spermatogenesis
In Vitro Techniques
Real-Time Polymerase Chain Reaction
Theoretical Models

Keywords

  • Human gonadotropins
  • Leydig cells
  • Prepubertal biomimetic testis
  • Sertoli cells

ASJC Scopus subject areas

  • Urology

Cite this

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. / Arato, Iva; Luca, Giovanni; Mancuso, Francesca; Bellucci, Catia; Lilli, Cinzia; Calvitti, Mario; Hansen, Barbara C.; Milardi, Domenico; Grande, Giuseppe; Calafiore, Riccardo.

In: Asian Journal of Andrology, Vol. 20, No. 2, 01.01.2018, p. 160-165.

Research output: Contribution to journalArticle

Arato, Iva ; Luca, Giovanni ; Mancuso, Francesca ; Bellucci, Catia ; Lilli, Cinzia ; Calvitti, Mario ; Hansen, Barbara C. ; Milardi, Domenico ; Grande, Giuseppe ; Calafiore, Riccardo. / 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. In: Asian Journal of Andrology. 2018 ; Vol. 20, No. 2. pp. 160-165.
@article{6cf94f2ebb064871867828da4696a3a8,
title = "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",
abstract = "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-M{\"u}llerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD 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.",
keywords = "Human gonadotropins, Leydig cells, Prepubertal biomimetic testis, Sertoli cells",
author = "Iva Arato and Giovanni Luca and Francesca Mancuso and Catia Bellucci and Cinzia Lilli and Mario Calvitti and Hansen, {Barbara C.} and Domenico Milardi and Giuseppe Grande and Riccardo Calafiore",
year = "2018",
month = "1",
day = "1",
doi = "10.4103/aja.aja_47_17",
language = "English",
volume = "20",
pages = "160--165",
journal = "Asian Journal of Andrology",
issn = "1008-682X",
publisher = "Nature Publishing Group",
number = "2",

}

TY - JOUR

T1 - An in vitro prototype of a porcine biomimetic testis-like cell culture system

T2 - A novel tool for the study of reassembled Sertoli and Leydig cells

AU - Arato, Iva

AU - Luca, Giovanni

AU - Mancuso, Francesca

AU - Bellucci, Catia

AU - Lilli, Cinzia

AU - Calvitti, Mario

AU - Hansen, Barbara C.

AU - Milardi, Domenico

AU - Grande, Giuseppe

AU - Calafiore, Riccardo

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD 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.

AB - 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-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD 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.

KW - Human gonadotropins

KW - Leydig cells

KW - Prepubertal biomimetic testis

KW - Sertoli cells

UR - http://www.scopus.com/inward/record.url?scp=85042722909&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042722909&partnerID=8YFLogxK

U2 - 10.4103/aja.aja_47_17

DO - 10.4103/aja.aja_47_17

M3 - Article

VL - 20

SP - 160

EP - 165

JO - Asian Journal of Andrology

JF - Asian Journal of Andrology

SN - 1008-682X

IS - 2

ER -