Formation of organotypic testicular organoids in microwell culture†.
Animals
Cell Count
Cell Culture Techniques
/ instrumentation
Child, Preschool
Diethylhexyl Phthalate
/ analogs & derivatives
Humans
Infant
Macaca mulatta
Male
Mice
Organoids
/ cytology
Spermatogenesis
/ drug effects
Spermatogonia
/ cytology
Swine
Testis
/ cytology
Tissue Scaffolds
/ chemistry
Tretinoin
/ pharmacology
organoid
testicular morphogenesis
Journal
Biology of reproduction
ISSN: 1529-7268
Titre abrégé: Biol Reprod
Pays: United States
ID NLM: 0207224
Informations de publication
Date de publication:
01 06 2019
01 06 2019
Historique:
received:
19
02
2019
accepted:
29
03
2019
pubmed:
31
3
2019
medline:
12
8
2020
entrez:
31
3
2019
Statut:
ppublish
Résumé
Three-dimensional (3D) organoids can serve as an in vitro platform to study cell-cell interactions, tissue development, and toxicology. Development of organoids with tissue architecture similar to testis in vivo has remained a challenge. Here, we present a microwell aggregation approach to establish multicellular 3D testicular organoids from pig, mouse, macaque, and human. The organoids consist of germ cells, Sertoli cells, Leydig cells, and peritubular myoid cells forming a distinct seminiferous epithelium and interstitial compartment separated by a basement membrane. Sertoli cells in the organoids express tight junction proteins claudin 11 and occludin. Germ cells in organoids showed an attenuated response to retinoic acid compared to germ cells in 2D culture indicating that the tissue architecture of the organoid modulates response to retinoic acid similar to in vivo. Germ cells maintaining physiological cell-cell interactions in organoids also had lower levels of autophagy indicating lower levels of cellular stress. When organoids were treated with mono(2-ethylhexyl) phthalate (MEHP), levels of germ cell autophagy increased in a dose-dependent manner, indicating the utility of the organoids for toxicity screening. Ablation of primary cilia on testicular somatic cells inhibited the formation of organoids demonstrating an application to screen for factors affecting testicular morphogenesis. Organoids can be generated from cryopreserved testis cells and preserved by vitrification. Taken together, the testicular organoid system recapitulates the 3D organization of the mammalian testis and provides an in vitro platform for studying germ cell function, testicular development, and drug toxicity in a cellular context representative of the testis in vivo.
Identifiants
pubmed: 30927418
pii: 5423876
doi: 10.1093/biolre/ioz053
pmc: PMC7302515
doi:
Substances chimiques
Tretinoin
5688UTC01R
Diethylhexyl Phthalate
C42K0PH13C
mono-(2-ethylhexyl)phthalate
FU2EWB60RT
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
1648-1660Subventions
Organisme : NICHD NIH HHS
ID : R01 HD091068
Pays : United States
Organisme : NIH HHS
ID : R01 OD016575
Pays : United States
Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction.
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