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Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism

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Title: Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism
Authors: Wang, Danyang, Hildorf, Simone, Ntemou, Elissavet, Mamsen, Linn Salto, Dong, Lihua, Pors, Susanne Elisabeth, Fedder, Jens, Clasen-Linde, Erik, Cortes, Dina, Thorup, Jørgen, Andersen, Claus Yding
Source: Wang, D, Hildorf, S, Ntemou, E, Mamsen, L S, Dong, L, Pors, S E, Fedder, J, Clasen-Linde, E, Cortes, D, Thorup, J & Andersen, C Y 2022, 'Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism', International Journal of Molecular Sciences, vol. 23, no. 14, 7975. https://doi.org/10.3390/ijms23147975
Publication Year: 2022
Collection: University of Southern Denmark: Research Output / Syddansk Universitet
Subject Terms: cryptorchidism, fertility preservation, human immature testicular tissue, infertility, organotypic culture, testicular tissue cryopreservation, Spermatogenesis/physiology, Humans, Cryptorchidism/metabolism, Testis/metabolism, Infant, Male, Sertoli Cells/metabolism, Spermatogonia/metabolism
Description: Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis.
Document Type: article in journal/newspaper
File Description: application/pdf
Language: English
DOI: 10.3390/ijms23147975
Availability: https://portal.findresearcher.sdu.dk/da/publications/4843d4e0-cd65-49b1-a875-a99b095a3026
https://doi.org/10.3390/ijms23147975
https://findresearcher.sdu.dk/ws/files/205576612/ijms_23_07975_v2.pdf
Rights: info:eu-repo/semantics/openAccess
Accession Number: edsbas.64EE3E60
Database: BASE
Description
Abstract:Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis.
DOI:10.3390/ijms23147975