Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model

García‐Domínguez, Ximo; Marco‐Jiménez, Francisco; Viudes-de-Castro, M.P.; Vicente, J.S.

doi:10.3791/58055

Cited by 22 publications

(32 citation statements)

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“…Embryo vitrification and warming steps were adapted from the highly efficient protocol developed previously to cryopreserve rabbit embryos by vitrification [ 23 , 24 ]. This protocol allows the survival of >80% of the thawed embryos, having generated thousands of descendants in our laboratory since its implementation [ 24 ]. Briefly, vitrification was achieved in two steps.…”

Section: Methodsmentioning

confidence: 99%

Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

García‐Domínguez

Marco‐Jiménez

Peñaranda

et al. 2020

Animals

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Nowadays, assisted reproductive technologies (ARTs) are considered valuable contributors to our past, but a future without their use is inconceivable. However, in recent years, several studies have evidenced a potential impact of ART on long-term development in mammal species. To date, the long-term follow-up data are still limited. So far, studies have mainly focused on in vitro fertilization or in vitro culture, with information from gametes/embryos cryopreservation field being practically missing. Herein, we report an approach to determine whether a vitrified embryo transfer procedure would have long-term consequences on the offspring. Using the rabbit as a model, we compared animals derived from vitrified-transferred embryos versus those naturally conceived, studying the growth performance, plus the weight throughout life, and the internal organs/tissues phenotype. The healthy status was assessed over the hematological and biochemical parameters in peripheral blood. Additionally, a comparative proteomic analysis was conducted in the liver tissue to investigate molecular cues related to vitrified embryo transfer in an adult tissue. After vitrified embryo transfer, birth weight was increased, and the growth performance was diminished in a sex-specific manner. In addition, vitrified-transferred animals showed significantly lower body, liver and heart weights in adulthood. Molecular analyses revealed that vitrified embryo transfer triggers reprogramming of the liver proteome. Functional analysis of the differentially expressed proteins showed changes in relation to oxidative phosphorylation and dysregulations in the zinc and lipid metabolism, which has been reported as possible causes of a disturbed growth pattern. Therefore, we conclude that vitrified embryo transfer is not a neutral procedure, and it incurs long-term effects in the offspring both at phenotypic and molecular levels. These results described a striking example of the developmental plasticity exhibited by the mammalian embryo.

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Section: Methodsmentioning

confidence: 99%

Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

García‐Domínguez

Marco‐Jiménez

Peñaranda

et al. 2020

Animals

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“…After storage in liquid nitrogen, embryos were warmed and vitrification solution was removed, loading the embryos into a solution containing DPBS and 0.33 M sucrose for 5 min, followed by one bath in a solution of DPBS for another 5 min before transfer. Immediately after warming, the embryos were evaluated morphologically, and only embryos without damage in mucin coat or pellucid zone were transferred by laparoscopy into the oviduct of synchronised recipient females from a maternal line (Line A [29]) following the procedure described by Besenfelder and Brem [30] and García-Domínguez et al [29]. The re-establishing of both populations to generate the 19th (R19V) and 37th (R37V) generation was described by Marco-Jiménez et al [26].…”

Section: Embryo Vitrification and Transfer To Population Rederivationmentioning

confidence: 99%

“…Five-hundred-and-fifty-seven embryos were used from donors belonging to R18 and R36 generations. Vitrification and transfer were described elsewhere [26,28,29]. Briefly, the vitrification was carried out in two steps at room temperature (approximately 20-22 • C).…”

Section: Embryo Vitrification and Transfer To Population Rederivationmentioning

confidence: 99%

Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations

Juarez

Marco‐Jiménez

Lavara

et al. 2020

Animals

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Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits such as birth and adult weights or prolificacy. The study aimed to evaluate the effect of a long-term selection for post-weaning average daily weight gain (ADG) over 37 generations with two rederived populations. Specifically, two coetaneous populations were derived from vitrified embryos with 18 generational intervals (R19 and R37), reducing or avoiding genetic drift and environmental and cryopreservation effects. After two generations of both rederived populations (R21 vs. R39 generations), all evaluated traits showed some progress as a result of the selection, the response being 0.113 g/day by generation. This response does not seem to affect the estimated Gompertz growth curve parameters in terms of the day, the weight at the inflexion point or the adult weight. Moreover, a sexual dimorphism favouring females was observed in this paternal line. Results demonstrated that the selection programme had improved ADG without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. Given the reduction in the cumulative reproductive performance and as a consequence in the selection pressure, or possibly/perhaps due to an unexpected effect, rederivation could be the cause of this weak selection response observed from generation 18 onwards.

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“…Vitrification was achieved in two steps, following the protocol adapted from previous studies [ 52 , 53 , 54 ]. Briefly, in the first step, embryos were placed for 2 min in a solution consisting of 10% ( v / v ) dimethyl sulfoxide (DMSO) and 10% ( v / v ) ethylene glycol (EG) in base medium.…”

Section: Methodsmentioning

confidence: 99%

“…Using this procedure, between 35 and 40 embryos were transferred to each pseudopregnant foster mother induced to ovulate 68–72 h before transfer. Both embryo vitrification and transfer processes used in this experiment were described in detail in Garcia-Dominguez et al [ 54 ].…”

Section: Methodsmentioning

confidence: 99%

Metabolomic Analysis Reveals Changes in Preimplantation Embryos Following Fresh or Vitrified Transfer

García‐Domínguez

Diretto

Frusciante

et al. 2020

IJMS

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Although assisted reproduction technologies (ARTs) are recognised as safe, and most of the offspring seem apparently healthy, there is clear evidence that ARTs are associated with changes in the embryo’s developmental trajectory, which incur physiological consequences during the prenatal and postnatal stages of life. The present study aimed to address the influence of early (day-3 embryos) embryo transfer and cryopreservation on embryo survival, size, and metabolome at the preimplantation stage (day-6 embryos). To this end, fresh-transferred (FT) and vitrified-transferred (VT) embryos were compared using naturally-conceived (NC) embryos as a control reference. The results show that as in vitro manipulation was increased (NC < FT < VT), both embryo survival rate (0.91 ± 0.02, 0.78 ± 0.05 and 0.63 ± 0.05, for NC, FT, and VT groups, respectively) and embryo size (3.21 ± 0.49 mm, 2.15 ± 0.51 mm, 1.76 ± 0.46 mm of diameter for NC, FT, and VT groups, respectively) were significantly decreased. Moreover, an unbiased metabolomics analysis showed overall down-accumulation in 40 metabolites among the three experimental groups, with embryo transfer and embryo cryopreservation procedures both exerting a cumulative effect. In this regard, targeted metabolomics findings revealed a significant reduction in some metabolites involved in metabolic pathways, such as the Krebs cycle, amino acids, unsaturated fatty acids, and arachidonic acid metabolisms. Altogether, these findings highlight a synergistic effect between the embryo transfer and vitrification procedures in preimplantation embryos. However, the ex vivo manipulation during embryo transfer seemed to be the major trigger of the embryonic changes, as the deviations added by the vitrification process were relatively smaller.

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Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model

Cited by 22 publications

References 0 publications

Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model

Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations

Metabolomic Analysis Reveals Changes in Preimplantation Embryos Following Fresh or Vitrified Transfer

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