» Articles » PMID: 24091557

In Vitro Sperm Production from Mouse Spermatogonial Stem Cell Lines Using an Organ Culture Method

Overview
Journal Nat Protoc
Specialties Biology
Pathology
Science
Date 2013 Oct 5
PMID 24091557
Citations 65
Authors
Affiliations
Soon will be listed here.
Abstract

The in vitro propagation of mouse spermatogonial stem cells (SSCs) became possible in 2003; these cultured SSCs were named germ-line stem (GS) cells. To date, however, it has not been possible to induce spermatogenesis from GS cells in vitro. Recently, we succeeded in producing functional sperm from primitive spermatogonia in explanted neonatal mouse testis tissues. Here we describe a protocol that can support spermatogenesis from GS cells up to sperm formation in vitro using an organ culture method. GS cells transplanted in the extracted testis form colonies in the tissue fragments and differentiate into sperm under the described in vitro organ culture conditions. It takes about 6 weeks to obtain sperm from GS cells. The sperm are viable, resulting in healthy offspring through micro-insemination. Thus, this protocol should be a valuable tool for the study of mammalian spermatogenesis.

Citing Articles

The Organotypic Culture of Mouse Seminiferous Tubules as a Reliable Methodology for the Study of Meiosis In Vitro.

Lopez-Jimenez P, Berenguer I, Perez-Moreno I, de Aledo J, Parra M, Page J Methods Mol Biol. 2024; 2818:147-160.

PMID: 39126472 DOI: 10.1007/978-1-0716-3906-1_9.


Generation of Meiotic Mouse Models Using CRISPR/Cas9 Technology.

Sanchez-Martin M, Sanchez-Saez F, Llano E, Pendas A Methods Mol Biol. 2024; 2818:93-112.

PMID: 39126469 DOI: 10.1007/978-1-0716-3906-1_6.


Recent Progress of Induced Spermatogenesis In Vitro.

Liu S, Wu J, Zhao X, Yu M, Taniguchi M, Bao H Int J Mol Sci. 2024; 25(15).

PMID: 39126092 PMC: 11313507. DOI: 10.3390/ijms25158524.


Targeting dysregulated phago-/auto-lysosomes in Sertoli cells to ameliorate late-onset hypogonadism.

Deng Z, Zhao L, Li S, Chen X, Ling X, Zheng J Nat Aging. 2024; 4(5):647-663.

PMID: 38649614 DOI: 10.1038/s43587-024-00614-2.


Male Infertility: New Developments, Current Challenges, and Future Directions.

Gul M, Russo G, Kandil H, Boitrelle F, Saleh R, Chung E World J Mens Health. 2024; 42(3):502-517.

PMID: 38164030 PMC: 11216957. DOI: 10.5534/wjmh.230232.


References
1.
Brinster R, Zimmermann J . Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A. 1994; 91(24):11298-302. PMC: 45218. DOI: 10.1073/pnas.91.24.11298. View

2.
Sato T, Katagiri K, Yokonishi T, Kubota Y, Inoue K, Ogonuki N . In vitro production of fertile sperm from murine spermatogonial stem cell lines. Nat Commun. 2011; 2:472. DOI: 10.1038/ncomms1478. View

3.
Gohbara A, Katagiri K, Sato T, Kubota Y, Kagechika H, Araki Y . In vitro murine spermatogenesis in an organ culture system. Biol Reprod. 2010; 83(2):261-7. DOI: 10.1095/biolreprod.110.083899. View

4.
Ogawa T, Ohmura M, Tamura Y, Kita K, Ohbo K, Suda T . Derivation and morphological characterization of mouse spermatogonial stem cell lines. Arch Histol Cytol. 2005; 67(4):297-306. DOI: 10.1679/aohc.67.297. View

5.
Kanatsu-Shinohara M, Ogonuki N, Inoue K, Miki H, Ogura A, Toyokuni S . Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol Reprod. 2003; 69(2):612-6. DOI: 10.1095/biolreprod.103.017012. View