Strains matter: Success of murine in vitro spermatogenesis is dependent on genetic background

Portela, Joana M D; Mulder, Callista L; Daalen, Saskia K.M. van; Winter-Korver, Cindy M de; Stukenborg, Jan-Bernd; Repping, Sjoerd; Pelt, Ans M.M. van

doi:10.1016/j.ydbio.2019.08.007

Cited by 17 publications

(18 citation statements)

References 33 publications

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“…However, lowering the temperature to 34 C did not prevent the observed block in in vitro meiotic divisions and haploid spermatid production. Moreover, in our experiments comparing two Sertoli cell lines and in testis tissue fragment cultures (Portela et al, 2019b), the somatic background seems to have a determinative effect on the germ cell developmental potential in vitro. Apart from the somatic supporting cells, the genetic background of the SSCs also determines how well they can be maintained in vitro (Kanatsu-Shinohara et al, 2003;Portela et al, 2019b).…”

Section: Discussionmentioning

confidence: 60%

“…Moreover, in our experiments comparing two Sertoli cell lines and in testis tissue fragment cultures (Portela et al, 2019b), the somatic background seems to have a determinative effect on the germ cell developmental potential in vitro. Apart from the somatic supporting cells, the genetic background of the SSCs also determines how well they can be maintained in vitro (Kanatsu-Shinohara et al, 2003;Portela et al, 2019b). Should any form of in vitro gametogenesis be considered as treatment for human infertility, the molecular mechanism that underlies these differences should be investigated.…”

Section: Discussionmentioning

confidence: 60%

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In Vitro Meiosis of Male Germline Stem Cells

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Section: Discussionmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 60%

In Vitro Meiosis of Male Germline Stem Cells

et al. 2020

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“…Soon later, the critical ingredients in KSR was discovered to be AlbuMAX, a purified serum albumin 3 . Because of this simple trick, the IVS with mouse testis tissue culture was repeated by several independent research groups and now becoming a standard procedure [4][5][6][7][8] . We continued working on the IVS to improve its efficiency by, for one thing, introducing microfluidic system.…”

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confidence: 99%

“…It was reported recently that mouse strain difference drastically influenced the efficiency of IVS with organ culture method6 . It would be possible therefore that the result of rat IVS differs in different strains.…”

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confidence: 99%

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

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In vitro spermatogenesis (IVS) using air–liquid interphase organ culture method is possible with mouse testis tissues. The same method, however, has been hardly applicable to animals other than mice, only producing no or limited progression of spermatogenesis. In the present study, we challenged IVS of rats with modifications of culture medium, by supplementing chemical substances, including hormones, antioxidants, and lysophospholipids. In addition, reducing oxygen tension by placing tissues in an incubator of lower oxygen concentration and/or applying silicone cover ceiling on top of the tissue were effective for improving the spermatogenic efficiency. Through these modifications of the culture condition, rat spermatogenesis up to round spermatids was maintained over 70 days in the cultured tissue. Present results demonstrated a significant progress in rat IVS, revealing conditions commonly favorable for mice and rats as well as finding rat-specific optimizations. This is an important step towards successful IVS in many animal species, including humans.

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“…It is important to highlight that different mouse strains have differences in physiology and results obtained herein could not give the same results for another mouse strain. A research group around Jan‐Bernd Stukenborg and Ans MM van Pelt showed that spermatogenic potential in vitro is dependent on specialized sites in the genome and therefore the organ culture conditions suboptimal for some strains of mice, like hybrid DBA2 strain with 75% C57BL/6J background (B6D2F2) 47 . The CD‐1 strain seems therefore adequate for this type of study, although other strains appear to give better results or to be more appropriate.…”

Section: Discussionmentioning

confidence: 99%

Activation of the cannabinoid receptor type 2 by the agonist JWH133 promotes the first wave of in vitro spermatogenesis

et al. 2020

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Background Oncological procedures have irreversible side effects on germ cells for childhood cancer survival boys. In vitro culture of prepubertal testicular tissue has been proposed to restore fertility; however, recent data on animal models showed that meiotic and post‐meiotic progression was impaired. Objectives As potential key inducers of the mitosis‐meiosis switch, type 2 cannabinoid receptor (CB2) has been proposed to play a central role in the meiotic entry of male germ cells. Herein, the in vitro first spermatogenesis wave in mice was used to understand the impact of CB2 activation on the differentiation of spermatogonia until elongated spermatids. Materials and methods A first set of cultured testicular explants of 6.5 days post‐partum (dpp) mice was performed to assess the impact of a range of JWH133 supplementation (10 nm, 100 nm, 1 µm, 10 µm). Then, the progressive development of germ cells at key timepoints of spermatogenesis was evaluated throughout (i) in vitro culture (day 2 [D2], D3, D6, D10, D18, and D30) coupled with (ii) in vivo counterparts (8.5, 9.5, 12.5, 16.5, 24.5, and 36.5 dpp). Results CB2 was detected at the plasma membrane of cells, and a successful completion of spermatogenesis was obtained in vitro. One day after the activation of CB2 by 1 μm of the agonist JWH133, percentage of zygotene spermatocyte I increased. Conclusion After 30 days of culture, (i) an enrichment of haploid germ cells detected by flow cytometry, (ii) a reduced necrotic area, and (iii) an increase in the density of post‐meiotic germ cells were observed. We showed that the activation of CB2 improves in vitro entry into meiosis and differentiation of spermatogonia, mimicking physiological meiotic transition.

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Strains matter: Success of murine in vitro spermatogenesis is dependent on genetic background

Cited by 17 publications

References 33 publications

In Vitro Meiosis of Male Germline Stem Cells

In Vitro Meiosis of Male Germline Stem Cells

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

Activation of the cannabinoid receptor type 2 by the agonist JWH133 promotes the first wave of in vitro spermatogenesis

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