Ultrastructural Morphometry of Bovine Blastocysts Produced In Vivo or In Vitro1

Crosier, Adrienne E.; Farin, P.W.; Dykstra, Michael J.; Alexander, Jacob; Farin, Charlotte E.

doi:10.1095/biolreprod64.5.1375

Cited by 150 publications

(117 citation statements)

References 47 publications

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“…Other authors have observed changes in the morphology of preimplantation embryos after culture. Crosier et al (2001) analyzed the ultrastructural morphometry of bovine blastocysts produced in vivo or in vitro using three different culture media and noted that in vitro blastocysts had a smaller nucleocytoplasmic ratio. They associated this finding with reduced quality of in vitro embryos.…”

Section: Discussionmentioning

confidence: 99%

Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos

Giritharan¹,

Talbi²,

Donjacour³

et al. 2007

Reproduction

142

127

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In vitro culture (IVC) of preimplantation mouse embryos is associated with changes in gene expression. It is however, not known if the method of fertilization affects the global pattern of gene expression. We compared gene expression and development of mouse blastocysts produced by in vitro fertilization (IVF) versus blastocysts fertilized in vivo and cultured in vitro from the zygote stage (IVC) versus control blastocysts flushed out of the uterus on post coital day 3.5. The global pattern of gene expression was assessed using the Affymetrix 430 2.0 chip. It appears that each method of fertilization has a unique pattern of gene expression and development. Embryos cultured in vitro had a reduction in the number of trophoblastic cells (IVF 33.5 cells, IVC 39.9 cells, and 49.6 cells in the in vivo group) and, to a lesser degree, of inner cell mass cells (12.8, 11.7, and 13.8 respectively). The inner cell mass nuclei were larger after culture in vitro (140 mm 2 , 113 mm 2 , and 86 mm 2 respectively). Although a high number of genes (1912) was statistically different in the IVF cohort when compared with the in vivo control embryos, the magnitude of the changes in gene expression were low and only a minority of genes (29 genes) was changed more than fourfold. Surprisingly, IVF embryos were different from IVC embryos (3058 genes were statistically different, but only three changed more than fourfold). Proliferation, apoptosis, and morphogenetic pathways are the most common pathways altered after IVC. Overall, IVF and embryo culture have a profound effect on gene expression pattern and phenotype of mouse preimplantation embryos.

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Section: Discussionmentioning

confidence: 99%

Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos

Giritharan¹,

Talbi²,

Donjacour³

et al. 2007

Reproduction

142

127

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“…It has been also reported that embryos developed in serum containing media have a higher proportion of TE cells compared with embryos produced in serum-free media [48]. TE cells are crucial for blastocoele re-expansion and maintenance after cryopreservation, and the higher lipid contents in TE than the ICM [26] could make TE cells particularly damaged during cryopreservation.…”

Section: Discussionmentioning

confidence: 99%

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

et al. 2008

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The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2 + 5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF) + 5% FCS, and SOF + 20 g L À1 bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system neither affects the embryo development up to Day 7 nor survival rates after vitrification. The culture of embryos in SOF + FCS and in Vero cells + B2 allowed obtaining more Day 6 and Day 7 blastocysts, and a higher % of Day 7 blastocysts vitrified than culture in SOF + BSA. Contrarily, on Day 8, more blastocysts were vitrified in SOF + BSA than in SOF + FCS. Blastocysts quality affected development after vitrification/warming, and Day 7 embryos showed higher survival rates than their Day 8 counterparts. Day 7 blastocysts produced in Vero cells or in SOF + BSA survived at higher rates than those produced in SOF + FCS at 24 and 48 h after warming. Embryo culture with BSA allows obtaining hatching rates after vitrification/warming higher than those obtained after coculture with Vero cells in B2 and FCS. Moreover, this system provides hatching rates from Day 8 blastocysts comparable to those obtained on Day 7 in Vero cells. Further studies, including embryo transfer to recipients, are needed to clarify factors affecting the freezability of in vitro produced bovine embryos. #

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“…This lower cryotolerance has been associated with the high lipid content present in the cytoplasm of these embryos (Abe et al, 2002;Mucci et al, 2006) and the decrease in the density of mature mitochondria compared to embryos produced in vivo (Crosier et al, 2001;Farin et al, 2004). Additionally, the most abundant lipids in the plasma membranes of cells (phosphatidylcholine and sphingomyelin) also have different profiles (Sudano et al, 2012b).…”

Section: Cryopreservation Of Bovine Embryosmentioning

confidence: 99%

Cryopreservation of in vitro-produced embryos: challenges for commercial implementation

Sanches¹,

Zangirólamo²,

Silva³

et al. 2017

Anim Reprod

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In the last several years, the high demand for embryo production has resulted in the need to study new methods to make the cryopreservation of bovine embryos produced in vitro more efficient. Despite the advantages offered by in vitro embryo production (IVEP), the major challenge to its greater dissemination is to improve embryonic survival after cryopreservation. Embryos that are produced in vitro are less resistant to cryopreservation compared to those produced in vivo, which is due to the higher accumulation of lipids in their cells, among other factors. In this context, changes in the culture conditions such as the addition of lipolytic chemical substances and the adjustment of fetal calf serum in the medium have been proposed to decrease the lipid amount within the embryos. Several years ago, vitrification allowed good results for in vitro produced (IVP) embryos because of its simplicity, speed and low cost. More recently, another technique applied to simplify the embryo post-thawing rehydration step in vivo, direct transfer (DT), is a strategy that has proven to be of interest in helping to overcome limitations to the cryopreservation of in vitro produced embryos. DT has been performed by commercial laboratories, ensuring good embryo viability after thawing. However, commercial and operational limitations prevent the large-scale use of these techniques. Thus, this review aims to discuss the use of strategies to improve the postcryopreservation survival capacity and the aspects to be overcome so that the cryopreservation of IVP embryos becomes a well-established and commercially applicable technique in addition to presenting new guidelines for embryo transfer (ET) programs from a better selection of recipients.

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Ultrastructural Morphometry of Bovine Blastocysts Produced In Vivo or In Vitro1

Cited by 150 publications

References 47 publications

Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos

Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

Cryopreservation of in vitro-produced embryos: challenges for commercial implementation

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