First birth of an animal from an extinct subspecies (Capra pyrenaica pyrenaica) by cloning

Folch, J.; Cocero, M.J.; Chesné, P.; Alabart, J. L.; Domínguez, Verónica; Cognie, Y.; Roche, A.; Fernández-Arias, A.; Martí, J.I.; Sánchez, P Nofre; Echegoyen, E.; Beckers, J. F.; Bonastre, A. Sánchez; Vignon, Xavier

doi:10.1016/j.theriogenology.2008.11.005

Cited by 147 publications

(102 citation statements)

References 21 publications

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“…Although androgenesis performed by using frozen sperm and γ-ray-inactivated xenogenic eggs can regenerate live fish, their survival rate is extremely low, and resulting offspring become nuclear-cytoplasmic hybrids (11). The loss of maternally inherited materials including mitochondrial DNA makes this method impractical, as it also does for the transfer of nuclei from cryopreserved somatic cells into xenogenic oocytes (12,13). In zebrafish, in vitro maturation of immature oocytes subsequent to cryopreservation is also impossible.…”

mentioning

confidence: 99%

Generation of functional eggs and sperm from cryopreserved whole testes

Lee

Iwasaki

Shikina

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

133

118

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The conservation of endangered fish is of critical importance. Cryobanking could provide an effective backup measure for use in conjunction with the conservation of natural populations; however, methodology for cryopreservation of fish eggs and embryos has not yet been developed. The present study established a methodology capable of deriving functional eggs and sperm from frozen type A spermatogonia (ASGs). Whole testes taken from rainbow trout were slowly frozen in a cryomedium, and the viability of ASGs within these testes did not decrease over a 728-d freezing period. Frozenthawed ASGs that were intraperitoneally transplanted into sterile triploid hatchlings migrated toward, and were incorporated into, recipient genital ridges. Transplantability of ASGs did not decrease after as much as 939 d of cryopreservation. Nearly half of triploid recipients produced functional eggs or sperm derived from the frozen ASGs and displayed high fecundity. Fertilization of resultant gametes resulted in the successful production of normal, frozen ASG-derived offspring. Feasibility and simplicity of this methodology will call for an immediate application for real conservation of endangered wild salmonids.fish genetic resources | testes cryopreservation | germ cell transplantation | stem cell | triploid trout B ecause of the environmental effects of human activities, a growing number of fish species have become threatened or are already extinct (1). Salmonid species are particularly vulnerable to habitat destruction and the effects of climate change as they occupy a unique ecological niche resulting from their longrange migration between freshwater and marine habitats (2, 3). Conservation strategies for these threatened fish must therefore be developed and implemented with all possible haste. Complicating the conservation of threatened salmonid populations are the risks associated with captive breeding and translocation of live fish, such as facility accidents, pathogen infections, genetic drift, and reduced fitness of individuals within natural habitats (4, 5).Cryobanking of fish gametes to semipermanently (6) store genetic resources could serve as an alternative approach to traditional conservation methods (7); however, past attempts at cryopreservation of fish embryos and mature oocytes have been unsuccessful (8), as a result of their large size, high yolk content, and low membrane permeability (9, 10). Although androgenesis performed by using frozen sperm and γ-ray-inactivated xenogenic eggs can regenerate live fish, their survival rate is extremely low, and resulting offspring become nuclear-cytoplasmic hybrids (11). The loss of maternally inherited materials including mitochondrial DNA makes this method impractical, as it also does for the transfer of nuclei from cryopreserved somatic cells into xenogenic oocytes (12, 13). In zebrafish, in vitro maturation of immature oocytes subsequent to cryopreservation is also impossible. Although techniques for cryopreservation of stage I and II oocytes (diameter, 90-350 μm) (14) and ...

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mentioning

confidence: 99%

Generation of functional eggs and sperm from cryopreserved whole testes

Lee

Iwasaki

Shikina

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

133

118

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“…Highly valuable deceased animals, either due to their genetic background or because they belonged to endangered species, might thus be recreated using nuclear transfer, if an effective and side effect-free somatic cell nuclear transfer was available. As a proof of principle, an extinct wild goat, the bucardo, has been cloned from frozen fibroblasts isolated from skin biopsies taken from the last survivor, a female that died in 1999 (Folch et al 2009). Certainly, there is a time limit for recreating deceased animals, which at the moment can be stored only using conventional systems.…”

Section: Discussionmentioning

confidence: 99%

Genome of non-living cells: trash or recycle?

Loi

Fulka²,

Hildebrand³

et al. 2011

Reproduction

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Reproductive technologies have been often used as a tool in research not strictly connected with developmental biology. In this study, we retrace the experimental routes that have led to the adoption of two reproductive technologies, ICSI and somatic cell nuclear transfer (SCNT), as biological assays to probe the 'functionality' of the genome from dead cells. The structural peculiarities of the spermatozoa nucleus, namely its lower water content and its compact chromatin structure, have made it the preferred cell for these experiments. The studies, primarily focused on mice, have demonstrated an unexpected stability of the spermatozoa nuclei, which retained the capacity to form pronuclei once injected into the oocytes even after severe denaturing agents like acid treatment and high-temperature exposure. These findings inspired further research culminating in the production of mice after ICSI of lyophilized spermatozoa. The demonstrated non-equivalence between cell vitality and nuclear vitality in spermatozoa prompted analogous studies on somatic cells. Somatic cells were treated with the same physical stress applied to spermatozoa and were injected into enucleated sheep oocytes. Despite the presumptive fragile nuclear structure, nuclei from non-viable cells (heat treated) directed early and post-implantation embryonic development on nuclear transfer, resulting in normal offspring. Recently, lyophilized somatic cells used for nuclear transfer have developed into normal embryos. In summary, ICSI and SCNT have been useful tools to prove that alternative strategies for storing banks of non-viable cells are realistic. Finally, the potential application of freeze-dried spermatozoa and cells is also discussed.

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“…Using cryopreserved tissue from the last known Pyrenean ibex, a Spanish group used somatic cell nuclear transfer (SCNT) to revive that extinct subspecies. Out of several hundred efforts, however, only one fetus survived to term, and it died minutes after birth from lung abnormalities ( 4). This example highlights two problems with SCNT: it is neither very safe nor effi cient and will only work if viable cell nuclei are available.…”

Section: Policyforummentioning

confidence: 99%