Successful Spermatogonial Stem Cells Transplantation within Pleuronectiformes: First Breakthrough at inter-family Level in Marine Fish

Zhou, Li; Liu, Qinghua; Yang, Jingkun; Xu, Shihong; Wu, Zhihao; Wang, Yanfeng; You, Feng; Song, Zongcheng; Li, Jun

doi:10.7150/ijbs.63266

Cited by 21 publications

(12 citation statements)

References 52 publications

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“…Similarly, goldfish recipients have also produced donor‐derived gametes from common carp (Franěk et al, 2021). Donor‐derived sperm production has also been reported in several xenogeneic transplantations between donor‐ and recipient‐species, such as rare Chinese minnow ( Gobiocypris rarus ) and zebrafish (Danio rerio ) (Zhang et al, 2021), and summer flounder ( Paralichthys dentatus ) or turbot ( Scophthalmus maximus ) and Japanese flounder ( P. olivaceus ) (Zhou et al, 2021).…”

Section: Discussionmentioning

confidence: 97%

Enrichment of spermatogonia by density gradient centrifugation for use as a donor of surrogate production of tiger puffer

Yoshikawa

Ino

Shigematsu

et al. 2022

Aquaculture Research

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A method of producing functional gametes from cryopreserved testicular germ cells using surrogate broodstock technology is a powerful tool for preserving the aquacultural genetic resources of the tiger puffer (Takifugu rubripes). Among the various testicular cells, only type A spermatogonia (ASGs) can colonize recipient gonads and shift to gametogenesis; therefore, ASG enrichment is an important factor for successful surrogate production. This study examined the possibility of ASG enrichment by density gradient centrifugation for surrogate production, considering its practical use. Through density gradient centrifugation with Percoll media, the highest distribution of ASGs estimated on the cell morphology and vasa expression was counted at a fraction of 20% Percoll layer. These fractionated testicular cells were transplanted intraperitoneally and successfully incorporated into the gonads of the recipient, grass puffer (T. alboplumbeus); however, no colonization was observed in the control recipients transplanted with unfractionated testicular cells. Moreover, these recipients matured normally and produced functional donor‐derived gametes from which tiger puffer offspring originated. Thus, the Percoll gradient centrifugation developed in this study enables the enrichment of ASGs from testicular cells, including many differentiated germ cells, and would be useful for efficiently managing broodstock with desirable genetic traits.

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Section: Discussionmentioning

confidence: 97%

Enrichment of spermatogonia by density gradient centrifugation for use as a donor of surrogate production of tiger puffer

Yoshikawa

Ino

Shigematsu

et al. 2022

Aquaculture Research

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“…Piracanjuba is an endangered species; therefore, the use of juveniles is unsuitable for the isolation of SSCs for cryopreservation or transplantation into adult recipients. In the absence of adults, however, testes from juveniles could be used in tissue cryopreservation and SSC isolation for transplantation in larvae (Marinović et al 2019;Zhou et al 2021).…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation and transplantation of spermatogonia stem cells in piracanjuba Brycon orbignyanus (Characiformes: Characidae), an endangered fish species

López,

Monzani,

Carvalho

et al. 2024

Preprint

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Piracanjuba (Brycon orbignyanus) is an endangered fish species from the Neotropical region. The establishment of a cryobank using spermatogonial stem cells (SSCs) and subsequent production of a germline chimera is thus a promising strategy for such species. In the present work, procedures for the isolation and cryopreservation of piracanjuba SSCs and subsequent transplantation into sterile recipients were established. The piracanjuba SSCs were obtained by Percoll density gradient centrifugation and differential plating. SSC fractions were evaluated by relative ddx4 expression, alkaline phosphatase activity, and light microscopy. SSC cryopreservation was performed using five cryoprotectants at three different concentrations. The mix of the cells from the 20% and 30% Percoll density gradients showed 58.35 ± 0.03% purity of SSCs. The purity of SSCs increased to 66.00 ± 0.01% after differential plating. The relative ddx4 expression was 3.5 times higher in cells from the Percoll density gradient centrifugation than in the gonad and cells after differential plating. Propanediol (1M) was the most effective cryoprotector evaluated (P = 1.000), showing 90.75 ± 1.85% cell viability. Freshly isolated and cryopreserved cells from the Percoll density gradient centrifugation were transplanted into a sterile male adult triploid hybrid with germ cell-less gonads. SSCs were observed in the germinal epithelium of the testes of recipients 20 days after transplantation. The results are promising for obtaining functional germline chimeras in Neotropical fish. Consequently, the procedures established here can be applied in future actions for the conservation and reconstitution of the piracanjuba in case of extinction.

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“…The turbot and Japanese ounder broodstock were raised in 30m 3 circular tanks with owing seawater and fed with frozen sh. The method of preparing triploids Japanese ounder (used as recipients) and feeding was conducted as previously described (Zhou et al, 2021). Brie y, the triploids recipients were produced by cold-shocking of freshly fertilized eggs at 0-2℃ for 45 min.…”

Section: Fish and Sample Preparationmentioning

confidence: 99%

Glial cell line-derived neurotrophic factor (GDNF) is essential for colonization and expansion of turbot (Scophthalmus maximus) spermatogonia stem cells in recipient and in vitro culture

Duan¹,

Du²,

Zhou³

et al. 2023

Preprint

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Germ cell transplantation (GCT) is a promising biotechnology producing donor-derived gametes in surrogate recipients. It plays a critical role of protecting endangered species, propagation of elite species with desired traits as well as long-term preservation of genetic resources in combination with cryopreservation. However, GCT often fails because of the low colonization rate even complete loss of donor cells in recipients. Glial cell line-derived neurotrophic factor (GDNF) is necessary for self-renew of mammals spermatogonia stem cells (SSCs), and it also contributes to the proliferation of SSCs in vitro culture in some animals. In turbot (Scophthalmus maximus), we found that the expressions of gdnf and gfrα1a were predominantly observed in spermatogonia rather than somatic cells, which differed from their expression patterns in mammals. The efficiency of exogenous spermatogonia transplantation in Japanese flounder (Paralichthys olivaceus) larvae could be substantially enhanced by incubating donor cells from turbot with 100ng/ml GDNF prior to transplantation, resulting in a notable rise in colonization rate from 33%-50% to 61.5%. Besides, the addition of 20ng/ml GDNF in cell medium could also promote the proliferation of turbot SSCs in vitro. These results demonstrated the gdnf in turbot testis expression characteristics and suggested that addition of GNDF might be an effective way to improve the GCT efficiency and promote the SSCs expansion in vitro culture.

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Successful Spermatogonial Stem Cells Transplantation within Pleuronectiformes: First Breakthrough at inter-family Level in Marine Fish

Cited by 21 publications

References 52 publications

Enrichment of spermatogonia by density gradient centrifugation for use as a donor of surrogate production of tiger puffer

Enrichment of spermatogonia by density gradient centrifugation for use as a donor of surrogate production of tiger puffer

Cryopreservation and transplantation of spermatogonia stem cells in piracanjuba Brycon orbignyanus (Characiformes: Characidae), an endangered fish species

Glial cell line-derived neurotrophic factor (GDNF) is essential for colonization and expansion of turbot (Scophthalmus maximus) spermatogonia stem cells in recipient and in vitro culture

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