Rapid cooling of rabbit embryos in a synthetic medium

Teixeira, Magda; Commin, Loris; Gavin-Plagne, Lucie; Bruyère, Pierre; Buff, Samuel; Joly, Thierry

doi:10.1016/j.cryobiol.2018.07.006

Cited by 5 publications

(4 citation statements)

References 67 publications

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“…This synthetic medium induced acceptable viability rates, growth recoveries and pluripotency gene expression when supplemented with 4–5% cryoprotectant. These data are consistent with our previous studies on embryo cryopreservation [ 38 , 39 , 44 ] and with other studies which showed that FBS could be replaced by a commercial synthetic media containing a high molecular weight polymer when freezing human or mouse PSCs [ 52 , 53 , 54 , 55 ]. In our study, we compared CRYO3, a synthetic commercial product accredited for clinical purposes, with CryoStor ® CS10, a widely used stem cell freezing medium of unknown composition.…”

Section: Discussionsupporting

confidence: 93%

“…To avoid the risks associated with serum, home-made and commercial freezing media has been supplemented with natural macromolecules, such as soybean lecithin [ 33 ] and silk sericin [ 34 ], or synthetic macromolecules, such as liposomes [ 35 ], polysaccharides [ 36 ], and polyvinylpyrrolidones [ 37 ], with varying degrees of success depending on the cryopreserved cells. Previous studies have already demonstrated the effectiveness of one type of commercial chemically defined media, STEM ALPHA.CRYO3 (hereinafter referred to as CRYO3), in the cryopreservation of rabbit embryos [ 38 , 39 ]. CRYO3 is a patented medium that lacks serum, protein, and dextran and is manufactured according to good manufacturing practice, in accordance with directive 2001/83/EC.…”

Section: Introductionmentioning

confidence: 99%

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Insights into Species Preservation: Cryobanking of Rabbit Somatic and Pluripotent Stem Cells

Gavin-Plagne

Perold

Osteil

et al. 2020

IJMS

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Induced pluripotent stem cells (iPSCs) are obtained by genetically reprogramming adult somatic cells via the overexpression of specific pluripotent genes. The resulting cells possess the same differentiation properties as blastocyst-stage embryonic stem cells (ESCs) and can be used to produce new individuals by embryonic complementation, nuclear transfer cloning, or in vitro fertilization after differentiation into male or female gametes. Therefore, iPSCs are highly valuable for preserving biodiversity and, together with somatic cells, can enlarge the pool of reproductive samples for cryobanking. In this study, we subjected rabbit iPSCs (rbiPSCs) and rabbit ear tissues to several cryopreservation conditions with the aim of defining safe and non-toxic slow-freezing protocols. We compared a commercial synthetic medium (STEM ALPHA.CRYO3) with a biological medium based on fetal bovine serum (FBS) together with low (0–5%) and high (10%) concentrations of dimethyl sulfoxide (DMSO). Our data demonstrated the efficacy of a CRYO3-based medium containing 4% DMSO for the cryopreservation of skin tissues and rbiPSCs. Specifically, this medium provided similar or even better biological results than the commonly used freezing medium composed of FBS and 10% DMSO. The results of this study therefore represent an encouraging first step towards the use of iPSCs for species preservation.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Insights into Species Preservation: Cryobanking of Rabbit Somatic and Pluripotent Stem Cells

Gavin-Plagne

Perold

Osteil

et al. 2020

IJMS

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“…Rabbit embryos were produced by ovarian stimulation. Sexually mature New Zealand white rabbits were injected with follicle-stimulating hormone and gonadotropin-releasing hormone, followed by artificial insemination or breeding, as previously described (Teixeira et al 2018). Eight-cell-stage embryos (E1.5) were flushed from explanted oviducts 36–40 h after insemination and cultured in a 1:1:1 mixture of RPMI 1640 medium, DMEM, and Ham’s F10 (RDH medium; Thermo Fisher Scientific) at 38°C in 5% CO 2 until cell microinjection.…”

Section: Methodsmentioning

confidence: 99%

Generation of Systemic Chimeras via Rabbit Induced Pluripotent Stem Cells Reprogrammed with KLF2, ERAS, and PRMT6

Perold,

Pham,

Pijoff

et al. 2024

Preprint

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Little is known about the molecular underpinnings of pluripotent stem cells' (PSCs) ability to colonize the epiblast of preimplantation embryos and generate chimeras. In our study, using rabbit PSCs as a model system, we conducted unbiased screening of a cDNA library that encodes a panel of 36 pluripotency factors. From this screening, we identified KLF2, ERAS and PRMT6, whose overexpression confers the ability for self-renewal in a KOSR/FGF2-free culture medium supplemented with LIF, activin A, PKC and WNT inhibitors. The reprogrammed cells acquired transcriptomic and epigenetic features of naive pluripotency, including the reactivation of the 2nd X-chromosome. Leveraging these PSC lines, we determined the transcriptomic signature of embryonic colonization-competence, demonstrating transcriptional repression of genes involved in MAPK, WNT, HIPPO, and EPH signaling pathways, alongside the activation of genes involved in amino-acid metabolism, NF-kB signaling, and p53 pathway. Remarkably, a subset of reprogrammed cells, expressing CD75 at a high level, gained the ability to produce chimeric fetuses with a high contribution from PSCs in all lineages.

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“…All procedures in rabbits were approved by the French ethics committee CELYNE (approval number APAFIS#6438 and APAFIS #2180-2015112615371038v2), and COMETHEA n°45, registered under n°12/107 and n°15/59. Sexually mature New Zealand white rabbits were injected with follicle-stimulating hormone and gonadotropin-releasing hormone, followed by artificial insemination as previously described (Teixeira et al, 2018). Embryos were flushed from explanted oviducts 65–159 h after insemination.…”

Section: Methodsmentioning

confidence: 99%

Major transcriptomic, epigenetic and metabolic changes underly the pluripotency continuum in rabbit preimplantation embryos

Bouchereau

Jouneau

Archilla

et al. 2021

Preprint

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Despite the growing interest in the rabbit model for developmental and stem cell biology, the characterization of embryos at the molecular level is still poorly documented. We conducted a transcriptome analysis of rabbit pre-implantation embryos from E2.7 (morula stage) to E6.6 (early primitive streak stage) using bulk and single-cell RNA-sequencing, and single-cell Biomark qPCR. In parallel, we studied oxidative phosphorylation and glycolysis and analyzed active and repressive epigenetic modifications during blastocyst formation and expansion. We generated a transcriptomic, epigenetic, and metabolic map of the pluripotency continuum in rabbit preimplantation embryos and identified novel markers of naive pluripotency that might be instrumental for deriving naive pluripotent stem cell lines. Although the rabbit is evolutionarily closer to mice than to primates, we found that the transcriptome of rabbit epiblast cells shares common features with that of humans and non-human primates.

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Rapid cooling of rabbit embryos in a synthetic medium

Cited by 5 publications

References 67 publications

Insights into Species Preservation: Cryobanking of Rabbit Somatic and Pluripotent Stem Cells

Insights into Species Preservation: Cryobanking of Rabbit Somatic and Pluripotent Stem Cells

Generation of Systemic Chimeras via Rabbit Induced Pluripotent Stem Cells Reprogrammed with KLF2, ERAS, and PRMT6

Major transcriptomic, epigenetic and metabolic changes underly the pluripotency continuum in rabbit preimplantation embryos

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