Phospholipid composition and resistance to vitrification of in vivo blastocyst of a Brazilian naturalized porcine breed

Sprícigo, J. F. W.; Leme, L. O.; Guimarães, A. L. S.; Neto, JP Costa; Silva, P.C.P.; Moreira, N. H.; Pivato, I.; Silva, B.D.M.; Ramos, A. F.; Dode, M. A. N.

doi:10.1590/1678-4162-10249

Cited by 4 publications

(4 citation statements)

References 39 publications

(49 reference statements)

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“…We hypothesized that embryonic gene expression alterations necessary to overcome the vitrification damages and allowing embryo recovery are reflected in the embryos that successfully survived the VIT procedure and they could be used to evaluate the efficiency of the vitrification procedure. Mostly, the evaluation of the success of vitrification protocols is based on in vitro survival rates, which implies that an in vitro culture step is performed to evaluate the restructuration of the embryo and its quality after vitrification/warming [ 39 – 41 ]. This makes the analysis of specific effects of VIT on embryos very difficult and involves important limitations: 1) If the VIT embryos are used for RNA–seq analysis directly after warming, these embryos may include an unknown proportion of embryos which would not recover (did not survive VIT) and recovering ones, affecting the gene expression analysis; 2) If the VIT is followed by 24 h IVC to allow the embryos to recover, the challenge is to distinguish VIT from IVC effects.…”

Section: Discussionmentioning

confidence: 99%

“…Here, we used a more extensive transcriptomic approach by using RNA–sequencing to analyze molecular alterations induced by vitrification. Furthermore, most studies analyzing the effect of vitrification on the embryo gene expression used vitrified/warmed embryos that are cultured in vitro for a variable period of time allowing them to recover and re–expand (6 h, 24 h, 48 h) [ 39 – 41 ], avoiding to include dead embryos or embryos that are not completely recovered in the gene expression analysis. This leads to the analysis of the cumulative effect of VIT and IVC, while the specific alterations induced by VIT remain unknown.…”

Section: Introductionmentioning

confidence: 99%

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Unveiling how vitrification affects the porcine blastocyst: clues from a transcriptomic study

Almiñana

Dubuisson

Bauersachs

et al. 2022

J Animal Sci Biotechnol

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Background Currently, there is a high demand for efficient pig embryo cryopreservation procedures in the porcine industry as well as for genetic diversity preservation and research purposes. To date, vitrification (VIT) is the most efficient method for pig embryo cryopreservation. Despite a high number of embryos survives in vitro after vitrification/warming procedures, the in vivo embryo survival rates after embryo transfer are variable among laboratories. So far, most studies have focused on cryoprotective agents and devices, while the VIT effects on porcine embryonic gene expression remained unclear. The few studies performed were based on vitrified/warmed embryos that were cultured in vitro (IVC) to allow them to re–expand. Thus, the specific alterations of VIT, IVC, and the cumulative effect of both remained unknown. To unveil the VIT-specific embryonic alterations, gene expression in VIT versus (vs.) IVC embryos was analyzed. Additionally, changes derived from both VIT and IVC vs. control embryos (CO) were analyzed to confirm the VIT embryonic alterations. Three groups of in vivo embryos at the blastocyst stage were analyzed by RNA–sequencing: (1) VIT embryos (vitrified/warmed and cultured in vitro), (2) IVC embryos and (3) CO embryos. Results RNA–sequencing revealed three clearly different mRNA profiles for VIT, IVC and CO embryos. Comparative analysis of mRNA profiles between VIT and IVC identified 321, differentially expressed genes (DEG) (FDR < 0.006). In VIT vs. CO and IVC vs. CO, 1901 and 1519 DEG were found, respectively, with an overlap of 1045 genes. VIT-specific functional alterations were associated to response to osmotic stress, response to hormones, and developmental growth. While alterations in response to hypoxia and mitophagy were related to the sum of VIT and IVC effects. Conclusions Our findings revealed new insights into the VIT procedure-specific alterations of embryonic gene expression by first comparing differences in VIT vs. IVC embryos and second by an integrative transcriptome analysis including in vivo control embryos. The identified VIT alterations might reflect the transcriptional signature of the embryo cryodamage but also the embryo healing process overcoming the VIT impacts. Selected validated genes were pointed as potential biomarkers that may help to improve vitrification.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Unveiling how vitrification affects the porcine blastocyst: clues from a transcriptomic study

Almiñana

Dubuisson

Bauersachs

et al. 2022

J Animal Sci Biotechnol

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“…Of the 20 potentially relevant articles, one was an in vitro experimental study [ 46 ]; three articles were excluded because of the wrong population (cumulus cells [ 47 ] or older fetus [ 48 , 49 ]); and 10 published either no MSI results, only MS data [ 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 ], or only proteomics of the embryos [ 59 ]. One abstract was also excluded because no data extraction could be performed [ 60 ].…”

Section: Resultsmentioning

confidence: 99%

Lipid Changes in the Peri-Implantation Period with Mass Spectrometry Imaging: A Systematic Review

Gitta

Márk

Szentpéteri

et al. 2023

Life

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Mass spectrometry imaging is a sensitive method for detecting molecules in tissues in their native form. Lipids mainly act as energy stores and membrane constituents, but they also play a role in lipid signaling. Previous studies have suggested an important role of lipids in implantation; therefore, our aim was to investigate the lipid changes during this period based on the available literature. The systematic literature search was performed on Ovid MEDLINE, Cochrane Library, Embase, and LILACS. We included studies about lipid changes in the early embryonal stage of healthy mammalian development published as mass spectrometry imaging. The search retrieved 917 articles without duplicates, and five articles were included in the narrative synthesis of the results. Two articles found a different spatial distribution of lipids in the early bovine embryo and receptive uterus. Three articles investigated lipids in mice in the peri-implantation period and found a different spatial distribution of several glycerophospholipids in both embryonic and maternal tissues. Although only five studies from three different research groups were included in this systematic review, it is clear that the spatial distribution of lipids is diverse in different tissues and their distribution varies from day to day. This may be a key factor in successful implantation, but further studies are needed to elucidate the exact mechanism.

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“…Despite this technical advantage for polytocous species, very little information is available on the number of embryos that can be successfully vitrified at one time using the Cryotop R system. In porcine species, although some studies have reported the vitrification of five to six in vivo-derived blastocysts loaded onto the tip of a Cryotop R device (23)(24)(25), to the best of our knowledge, the simultaneous vitrification of a greater number of morulae and blastocysts using this system has not been investigated.…”

Section: Introductionmentioning

confidence: 99%

The Open Cryotop System Is Effective for the Simultaneous Vitrification of a Large Number of Porcine Embryos at Different Developmental Stages

Gonzalez-Plaza

Cambra²,

Parrilla³

et al. 2022

Front. Vet. Sci.

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The Superfine Open Pulled Straw (SOPS) system is the most commonly used method for vitrification of pig embryos. However, this system only allows the vitrification of four to seven embryos per straw. In this study, we investigated the effectiveness of the open (OC) and closed (CC) Cryotop® systems to simultaneously vitrify a larger number of porcine embryos. Morulae, early blastocysts and full blastocysts were vitrified with the open Cryotop® (n = 250; 20 embryos per device) system, the closed Cryotop® (n = 158; 20 embryos per device) system and the traditional superfine open pulled straw (SOPS; n = 241; 4–7 embryos per straw) method. Fresh embryos from each developmental stage constituted the control group (n = 132). Data expressed as percentages were compared with the Fisher's exact test. The Kruskal-Wallis test was used to analyze the effect of the different vitrification systems on the embryo quality parameters and two-by-two comparisons were accomplished with the Mann-Whitney U test. Differences were considered statistically significant when p < 0.05. Vitrified and control embryos were incubated for 24 h and examined for viability and quality. At the warming step, the embryo recovery rate for the CC system was 51%, while all embryos were recovered when using OC and SOPS. There were no differences between the vitrification and control groups in the postwarming viability of full blastocysts. In contrast, morulae and early blastocysts that were vitrified-warmed with the SOPS system had lower viability (p < 0.01) compared to those from the OC, CC and control groups. The embryonic viability was similar between the OC and control groups, regardless of the developmental stage considered. Moreover, the embryos from the OC group had comparable total cell number and cells from the inner cell mass and apoptotic index than the controls. In conclusion, the OC system is suitable for the simultaneous vitrification of 20 porcine embryos at different developmental stages and provides comparable viability and quality results to fresh embryos subjected to 24 h of in vitro culture.

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Phospholipid composition and resistance to vitrification of in vivo blastocyst of a Brazilian naturalized porcine breed

Cited by 4 publications

References 39 publications

Unveiling how vitrification affects the porcine blastocyst: clues from a transcriptomic study

Unveiling how vitrification affects the porcine blastocyst: clues from a transcriptomic study

Lipid Changes in the Peri-Implantation Period with Mass Spectrometry Imaging: A Systematic Review

The Open Cryotop System Is Effective for the Simultaneous Vitrification of a Large Number of Porcine Embryos at Different Developmental Stages

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