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Actions of FGF2, LIF, and IGF1 on Embryo Survival and Conceptus Elongation following Slow-rate Freezing

ABSTRACT: Actions of FGF2, LIF, and IGF1 on Embryo Survival and Conceptus Elongation following Slow-rate Freezing

PROVIDER: PRJNA888397 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21845905_1.fastq.gz	Fastqsanger.gz
	SRR21845905_2.fastq.gz	Fastqsanger.gz
	SRR21845906_1.fastq.gz	Fastqsanger.gz
	SRR21845906_2.fastq.gz	Fastqsanger.gz
	SRR21845907_1.fastq.gz	Fastqsanger.gz

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Actions of FGF2, LIF, and IGF1 on Embryo Survival and Conceptus Elongation following Slow-rate Freezing

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Project description:In vitro embryo production systems are limited by their inability to consistently produce embryos with the competency to develop to the blastocyst stage, survive cryopreservation, and establish a pregnancy. Previous work identified a combination of three cytokines [fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1)], called FLI, that we hypothesize improve preimplantation development of bovine embryos in vitro. To test this hypothesis, FLI was supplemented into oocyte maturation or embryo culture medium. Embryos were produced in vitro using abattoir-derived oocytes and fertilized with sperm from a single bull known to have high fertility. After an 18-20 h fertilization period, putative zygotes were cultured in synthetic oviductal fluid (SOF) for 8 days. The addition of FLI to the oocyte maturation medium increased (P < 0.05) the dissociation of transzonal projections at 12, 18, and 24 h of maturation, as well as, the proportion of oocytes that reached the metaphase II stage of meiosis. Additionally, lipid content was decreased (P < 0.05) in the blastocyst stage embryo. The addition of FLI during the culture period increased development to the blastocyst stage, cytoskeleton integrity, and survival following slow freezing, as well as, decreased post thaw cell apoptosis (P < 0.05). In conclusion, the supplementation of these cytokines in vitro has the potential to alleviate some of the challenges associated with the cryo-survival of in vitro produced bovine embryos through improving embryo development and embryo quality.

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Impact of media supplements FGF2, LIF and IGF1 on the genome activity of porcine embryos produced in vitro.

Project description:In this article, we focused on the impact of precisely chemically modified FLI maturation medium enriched with fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), insulin-like growth factor 1 (IGF1), and polyvinyl alcohol (PVA) and its potential to improve the efficiency of in vitro production of porcine embryos. We hypothesized that enhancing the composition of the maturation medium could result in an elevated production of embryos in vitro and can affect EGA. FLI medium resulted in a significantly higher rate of oocyte blastocyst maturation and formation compared to the control DMEM medium. In addition, immunocytochemical labelling confirmed the detection of UBF in 4-cell FLI parthenogenic embryos, suggesting similarities with natural embryo development. Through RNAseq analysis, upregulated genes present in 4-cell FLI embryos were found to play key roles in important biological processes such as cell proliferation, cell differentiation, and transcriptional regulation. Based on our findings, we demonstrated the positive influence of FLI medium in the evaluation of in vitro embryo production, EGA detection, transcriptomic and proteomic profile, which was confirmed by the positive activation of the embryonal genome in the 4-cell stage of parthenogenetically activated embryos.

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IGF1-mediated human embryonic stem cell self-renewal recapitulates the embryonic niche

Project description:Our understanding of signalling pathways that regulate human pluripotent epiblast cells is limited, despite their fundamental biological and clinical importance. In this study, we mined human embryo transcriptomics datasets, and identified expression of transcripts for the insulin and insulin growth factor 1 (IGF1) receptors, along with IGF1 ligand. Human embryos cultured in medium supplemented with IGF1 showed an increase in the proportion of NANOG-expressing pluripotent epiblast progenitor cells. Consequently, we generated a minimal chemically-defined culture medium informed by the signalling environment of the human embryo. We derived several human embryonic stem cell (hESC) lines and reprogrammed fibroblasts to induced pluripotent stem cells (iPSCs) in these conditions. Transcriptome and protein expression analyses confirmed that these hESCs and iPSCs expressed pluripotency-associated genes. Moreover, hESCs and iPSCs retained high viability, the ability to undergo spontaneous and directed differentiation, and a normal karyotype. By contrast, fibroblast growth factor 2 (FGF2) was detrimental to epiblast development and dispensable for hESC cultures maintained in exogenous insulin or IGF1. This is the first example of how integrating signalling insights from human blastocysts can be used to define hESC culture conditions that more closely recapitulate the embryonic niche.

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