Transcriptomic difference in bovine blastocysts following vitrification and slow freezing at morula stage

Gupta, Amitava Sen; Singh, Jaswant; Dufort, Isabelle; Robert, Claude; Dias, F. C. F.; Anzar, M.

doi:10.1371/journal.pone.0187268

Cited by 34 publications

(28 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Knowledge so far indicates that suboptimal conditions during ART may induce embryonic stress responses and affect embryo viability [ 10 , 11 ]. In addition, cryopreservation is known for its harmful impacts on embryonic health, placing embryos at risk from a variety of types of cryoinjury during temperature and phase transitions [ 18 , 27 ]. Such survival impairment is in agreement with previous studies performed in rabbits, where manipulations of either fresh or vitrified embryos resulted in negative developmental effects both under in vitro conditions and in vivo [ 22 , 23 , 26 , 28 , 29 , 30 ].…”

Section: Discussionmentioning

confidence: 99%

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

García‐Domínguez

Diretto

Frusciante

et al. 2020

IJMS

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 99%

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

García‐Domínguez

Diretto

Frusciante

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…We also know that, vitri cation negatively impacts porcine embryo quality, augmenting levels of apoptosis [19,20] or impairing ultrastructure [21]. Vitri cation has been found to cause modi cations in the embryo gene expression in several species [22][23][24][25][26][27]; however, the consequences of vitri cation on gene expression of mammalian embryos are poorly known.…”

Section: Introductionmentioning

confidence: 99%

“…General transcriptome experiments of vitri ed-warmed have been done only in bovine. Vitri cation of in vitro-produced (IVP) bovine embryos caused overexpression of apoptosis-related genes [25] and the repression of genes involved in cell differentiation, cell adhesion and metabolism of lipids [28]. It has been also demonstrated that vitri cation modi ed expression of genes related to stress response [29] in both IVP and in vivo-derived bovine blastocysts.…”

Section: Introductionmentioning

confidence: 99%

Vitrification of in Vivo-derived Porcine Morulae and Blastocysts Alters Metabolic and Stress Response Pathway Alterations

Cuello¹,

Martínez²,

Cambra³

et al. 2020

Preprint

View full text Add to dashboard Cite

BackgroundDespite reported promising farrowing rates after non-surgical and surgical transfers of vitrified in vivo-derived porcine morulae and blastocysts (range: 70-75%), their pregnancy loss is yet higher (10-20%) than it is for fresh embryos (< 2.5%). The present study was designed to investigate whether vitrification would affect the transcriptome of porcine morulae and blastocysts, using microarrays and qRT-PCR validation. Material and methods and resultsMorulae and blastocysts were collected from weaned sows (n=13) by laparotomy at Day 6 (Day 0=start of estrus). Of each embryo category, 50 morulae and 50 blastocysts were vitrified (Treatment group) while fresh morulae (n=40) and blastocysts (n=40) acted as control group. After one week of storage, vitrified embryos were cultured in vitrofor 24 h after warming. Non-vitrified morulae (n=40) and blastocysts (n=40), cultured in vitro for 24 h were used as controls. After the in vitro culture period, embryo viability was morphologically assessed. A total of 30 viable embryos per group and embryonic stage(three pools of 10), were subjected to gene expressionanalysis byusing a microarray approach. A fold change cut-off of ±1.5 and a p-unadjusted <0.05 were used to distinguish differentially expressed genes (DEGs). The survival rates of vitrified/warmed morulae and blastocysts were similar to those of the control (nearly 100%, n.s.).A total of 233 DEGs were identified in vitrified morulae (38 upregulated and 195 downregulated) and 105 (112 upregulated and 93 downregulated) in vitrified blastocysts compared to the control group.Transcriptome DEG profiles were dependent on developmental stage, and GO enrichment analysis mainly related DEGs tobiological processes.The vitrification/warming impact on morulae was mostly repression of gene expression with the exception of metabolism-related pathways. In the case of blastocysts, we noted the activation of the cell cycle, cellular senescence andof signaling pathways for TFGβ, p53, FoxO and MAPK. Disruption ofmetabolic-related pathways in morulae and steroid biosynthesis and gap junctions in blastocysts could be related to impaired embryo quality and developmental potential, despite the rather high post-warming survival rates seen in vitro.ConclusionIn conclusion, vitrification modified the transcriptome of in vivo-derived porcine morulae and blastocysts, resulting in moderate gene expression changes, which may disturb subsequent embryo development and pregnancy after embryo transfer.

show abstract

“…Differentially expressed genes (DEGs) between frozen and vitrified (Larman et al, 2011;Stinshoff et al, 2011) and between fresh and vitrified (Leme et al, 2016) IVP embryos have already been described. According to Gupta et al (2017), vitrified morulae exhibited improved cryosurvival compared to those obtained using the slow freezing method. However, an altered transcriptional profile related to the impairment of implantation and placentation in the uterus was revealed in blastocysts derived from vitrified morulae (Gupta et al, 2017).…”

mentioning

confidence: 99%

“…According to Gupta et al (2017), vitrified morulae exhibited improved cryosurvival compared to those obtained using the slow freezing method. However, an altered transcriptional profile related to the impairment of implantation and placentation in the uterus was revealed in blastocysts derived from vitrified morulae (Gupta et al, 2017).…”

mentioning

confidence: 99%

Transcriptional profiling of embryo cryotolerance

Marsico

Caetano

Rodrigues³

et al. 2020

Molecular Reproduction Devel

View full text Add to dashboard Cite

The cryosurvival of embryos is a complex process involving dynamic and integrated morphological, functional, and molecular changes. Here, we evaluated the transcriptional profiling of bovine embryos possessing high and low cryotolerance (HC and LC, respectively) by assessing the resumption of development. Embryos were produced in vitro (N = 1137) and cryopreserved (N = 894). Blastocysts samples possessed pronounced group individualization at RNA sequencing. A total of 114 genes were differentially expressed, and 27 and 84 genes were upregulated in HC and LC, respectively. Among the over-represented biological functions, cellular growth and proliferation, cell death and survival, and organismal survival were predicted to be activated, while cellular movement and cell-to-cell signaling were predicted to be inhibited in HC embryos. Enriched canonical pathways and upstream regulators related to cellular proliferation and survival (HC), inflammatory processes, and cell death (LC) were predicted to represent two embryonic molecular profiles present during the resumption of development after cryopreservation. The marked contrast in transcriptional profiles between HC and LC strongly suggests the influence of embryonic competence after cryopreservation on its respective transcriptome and indicated that HC and LC presented two different molecular strategies to overcome cryopreservation-related stress and resume postcryopreservation development.

show abstract

Transcriptomic difference in bovine blastocysts following vitrification and slow freezing at morula stage

Cited by 34 publications

References 68 publications

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

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

Vitrification of in Vivo-derived Porcine Morulae and Blastocysts Alters Metabolic and Stress Response Pathway Alterations

Transcriptional profiling of embryo cryotolerance

Contact Info

Product

Resources

About