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See detailCryopreservation of chicken primordial germ cells by vitrification and slow-freezing: a comparative study
Tonus, Céline ULg; Connan, Delphine ULg; Waroux, Olivier ULg et al

in Theriogenology (2017), 88

In the present study, we compare a classical slow freezing method and an aseptic vitrification technique to cryopreserve a stable Primordial Gem Cells (PGCs) line issued from the Ardennaise chicken breed ... [more ▼]

In the present study, we compare a classical slow freezing method and an aseptic vitrification technique to cryopreserve a stable Primordial Gem Cells (PGCs) line issued from the Ardennaise chicken breed. Viability immediately after warming was close to 80% and did not differ between the two cryopreservation methods. Proliferation tended to be slower for both cryopreservation methods compared to controls, but the difference was significant only for vitrification. No difference was found between the two methods after flow cytometry analysis of SSEA-1 expression and RT-PCR on several factors related to PGCs phenotype. After one week in culture, all cryopreserved cells reached controls main morphological and expanding (viability/proliferation) features. However, slow freezing generated more unwanted cells clusters than vitrification. After injection of the PGCs into recipient embryos, vitrified PGCs showed a clear, yet not significant, tendency to colonize the gonad at a higher rate than slow frozen PGCs. Slow freezing in cryovials remains simple, inexpensive and less technically demanding than vitrification. Nevertheless, the intrinsic advantages of our aseptic vitrification method and the present study suggest that this should be considered as safer than classical slow freezing for cryopreserving chicken PGCs. [less ▲]

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See detailVitrification: Research and state-of-the-art in humans and animals
Ectors, Fabien ULg

Scientific conference (2016, December 09)

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See detailVITRICELL: new efficient method for cryopreserving cells by vitrification
Connan, Delphine ULg; Ectors, Fabien ULg; Antoine, Nadine ULg et al

Conference (2016, September)

Using stem and related cells for clinical purposes requires efficient and bio-safe handling. Cryopreservation is a mandatory key step of storage and transportation, during which cells undergo extreme ... [more ▼]

Using stem and related cells for clinical purposes requires efficient and bio-safe handling. Cryopreservation is a mandatory key step of storage and transportation, during which cells undergo extreme physical and chemical conditions prone to alter their viability as well as their biological properties. Conventional slow-freezing often results in poor survival rates mainly due to excessive cell dehydration and water crystallization. We have addressed this problem by developing a new cryopreservation method based on aseptic and automatable vitrification in sealed french straws. Furthermore, only bio-safe and chemically defined cryopreservation media are used. We have demonstrated that, despite additional constraints, our aseptic vitrification process is more efficient (recovery rates, morphology, pluripotency…) than conventional slow freezing for cryopreserving human pluripotent stem cells (hPSCs). These results have been confirmed on various sensitive stem cell-like lines and embryos from human and non-human species. VITRICELL will soon provide researchers and clinicians with its vitrification kits, allowing to upgrade the current yields and safety after cryopreservation of their high-value cells. [less ▲]

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See detailLong term-cultured and cryopreserved primordial germ cells from various chicken breeds retain high proliferative potential and gonadal colonisation competency
Tonus, Céline ULg; Cloquette, Karine; Ectors, Fabien ULg et al

in Reproduction, Fertility and Development (2016), 28(5), 628-639

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See detailEquine cadaver ligaments : A new promising source of stem cells
Shikh Al Sook, Mohamad Khir ULg; Gabriel, Annick ULg; Salouci, Moustafa et al

Scientific conference (2015, November 07)

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See detailEctopic Expression of Retrotransposon-Derived PEG11/RTL1 Contributes to the Callipyge Muscular Hypertrophy.
Xu, Xuewen; Ectors, Fabien ULg; Davis, Erica E. et al

in PloS one (2015), 10(10), 0140594

The callipyge phenotype is an ovine muscular hypertrophy characterized by polar overdominance: only heterozygous +Mat/CLPGPat animals receiving the CLPG mutation from their father express the phenotype ... [more ▼]

The callipyge phenotype is an ovine muscular hypertrophy characterized by polar overdominance: only heterozygous +Mat/CLPGPat animals receiving the CLPG mutation from their father express the phenotype. +Mat/CLPGPat animals are characterized by postnatal, ectopic expression of Delta-like 1 homologue (DLK1) and Paternally expressed gene 11/Retrotransposon-like 1 (PEG11/RTL1) proteins in skeletal muscle. We showed previously in transgenic mice that ectopic expression of DLK1 alone induces a muscular hypertrophy, hence demonstrating a role for DLK1 in determining the callipyge hypertrophy. We herein describe newly generated transgenic mice that ectopically express PEG11 in skeletal muscle, and show that they also exhibit a muscular hypertrophy phenotype. Our data suggest that both DLK1 and PEG11 act together in causing the muscular hypertrophy of callipyge sheep. [less ▲]

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See detailLong term culture, cryopreservation and genetic modification of chicken primordial germ cells
Tonus, Céline ULg; Garcia Gil, Francisco José ULg; Cloquette, Karine et al

Poster (2015, October 16)

Avian primordial germ cells (PGCs) are precursor of gametes and appear during early stages of embryonic development. Under appropriate culture conditions, these cells can keep their germ cells properties ... [more ▼]

Avian primordial germ cells (PGCs) are precursor of gametes and appear during early stages of embryonic development. Under appropriate culture conditions, these cells can keep their germ cells properties in vitro and are foreseen as promising tools for developing efficient avian genetic engineering and preservation of germplasm. We propose original methods that allow long term expansion, efficient cryopreservation and genetic modification of primary cultures of undifferentiated PGCs. PGCs are collected from embryonic blood during their migratory period and grown in cell-culture insert in the presence of feeder cells (BRL). This physically separated co-culture system along with selective culture medium promoted emergence, selection and proliferation of PGCs lines. Forty percent of blood samples gave rise to lines originating from three commercial layer and two Belgian endangered breeds. PGCs lines were characterized for the expression of the stem cells and PGCs marker SSEA-1 by FACS. RT-PCR confirmed expression of germ-line specific markers (CVH, CDH, DAZL), pluripotency markers (cPouV, cSox2, cNanog), telomerase and CXCR4 receptor. All lines were male although isolated from pooled male and female blood samples. Two cryopreservation methods were developed based upon slow-freezing and aseptic vitrification. Both have shown a similar effectiveness in allowing storage without phenotype drift. Stably expressing lines were obtained by Lipofectamine® mediated transfection of a GFP plasmid. PGCs were subsequently injected in recipient embryos. Persistence of exogenous PGCs in the developing gonad of recipient embryos confirmed that PGCs retain their gonadal colonisation ability, both after long-term culture and after cryopreservation. [less ▲]

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See detailIntracellular concentration of cryoprotectant during vitrification and slow-freezing cryopreservation procedures
Vanderzwalmen, Pierre ULg; Ectors, Fabien ULg; Wirleitner, Barbara et al

in Tucker, Mickael; Lieberman, Juergen (Eds.) Vitrification in assisted reproduction: second edition (2015)

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See detailVitrification of oocytes and embryos: Finally a recognized technique, but still a source of concern and debate
Vanderzwalmen, Pierre ULg; Zech, Nicolas; Ectors, Fabien ULg et al

in Tucker, Mickael; Lieberman, Juergen (Eds.) Vitrification in assisted reproduction: second edition (2015)

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See detailPhysiologie humaine
Sherwood, Lauralee; Ectors, Fabien ULg

Book published by De Boeck Supérieur - 3e édition française (2015)

"Apprendre et comprendre comment fonctionne l'organisme en 750 pages en couleurs, couvrant toutes les grandes fonctions physiologiques" Nouveautés et adaptations de la 3e édition Dans cette 3e édition ... [more ▼]

"Apprendre et comprendre comment fonctionne l'organisme en 750 pages en couleurs, couvrant toutes les grandes fonctions physiologiques" Nouveautés et adaptations de la 3e édition Dans cette 3e édition près de 90% des figures ont été retouchées. La plupart des images des cellules et des figures anatomiques sont nouvelles, elles sont plus réalistes et en perspective. Dans cette édition, des fiches de révision pour chaque chapitre ont été ajoutées à la fin du livre afin que les étudiants puissent réviser les points clés. Des problèmes de réflexion permettent aux étudiants d'analyser des cas cliniques et des symptômes à partir de leurs connaissances. Un contenu riche et pédagogique Tout est mis en oeuvre, dans ce livre, pour aider l'étudiant dans son apprentissage: des contenus riches, détaillés et actuels; de nombreuses aides et précisions à propos de la théorie ainsi qu'une structure claire et logique. De plus, la théorie est ponctuée d'analogies et de références à l'expérience quotidienne. Des schémas et illustrations de qualité Les schémas et illustrations sont proposés dans des couleurs nettes et de qualité. Les couleurs utilisées respectent celles observées dans la réalité pour une meilleure compréhension et intégration des informations. Un guide de révision Cet ouvrage a été pensé de manière à favoriser l'apprentissage de la physiologie et tout particulièrement l'homéostasie. Des notes cliniques viennent ponctuer le discours théorique, des exercices de révision sont disponibles à la fin de chaque chapitre et, à la fin du livre, se trouvent des compélements d'informations au sujet de chaque chapitre, nommés "cartes d'étude". [less ▲]

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See detailEquine cadaver ligaments : A new promising source of stem cells
Shikh Al Sook, Mohamad Khir ULg; Gabriel, Annick ULg; Salouci, Moustafa et al

Poster (2015)

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See detailVitrification, an efficient cryopreservation procedure but still rising some debates
Vanderzwalmen, Pierre; Ectors, Fabien ULg; Stecher, Astrid et al

in Current Trends in Clinical Embryology (2014), 1(1), 34-45

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See detail25KDa-Thiamine triphosphatase is essential for spermatozoid development in mice
Kohn, Grégory ULg; Ectors, Fabien ULg; Lakaye, Bernard ULg et al

Poster (2014, May)

For many years, our laboratory has been interested in thiamine triphosphate (ThTP), a vitamin B1 derivative whose metabolism and physiological role remain unclear. Regarding its production, we have shown ... [more ▼]

For many years, our laboratory has been interested in thiamine triphosphate (ThTP), a vitamin B1 derivative whose metabolism and physiological role remain unclear. Regarding its production, we have shown that in E. coli and in rat brain mitochondria, ThTP is synthesized from thiamine diphosphate and inorganic phosphate through a chemiosmotic mechanism involving the FoF1- ATP synthase [1, 2]. In mammalian cells, its concentration is maintained at a low level through hydrolysis by a very specific cytosolic 25-kDa thiamine triphosphatase (ThTPase) [3]. In order to gain insight in the role of ThTP and ThTPase in mammalian tissues, we decided to generate a mouse strain invalidated in 25kDa-ThTPase with the hope that these mice will accumulate ThTP in their tissues. We obtained genetically modified embryonic stem (ES) cells from the Knockout Mouse Project (KOMP) repository. In those cells, one of the 25kDa-ThTPase alleles was replaced by a construction containing the lacZ and the neomycin resistance genes. Those ES cells were microinjected in blastocysts and the chimeric blastocysts were injected in a mouse uterus to generate chimerae. However, when we bred those mice with wild type mice, the construction was never transmitted to the pups. To explain this result, we selected those chimerae that presented a sex-reversal. In those mice, all the spermatozoids derive from the injected embryonic stem cells, so that half of the spermatozoids are expected to harbor the construction. However, after qPCR analysis, we observed that the spermatozoids with the construction were outnumbered by a factor of thousand. These results strongly suggest that the 25kDa-ThTPase is required for spermatozoid development. 1. Gangolf, M., Wins, P., Thiry, M., El Moualij, B. & Bettendorff, L. (2010) J. Biol. Chem. 285, 583-94. 2. Gigliobianco, T., Gangolf, M., Lakaye, B., Pirson, B., von Ballmoos, C., Wins, P. & Bettendorff, L. (2013) Scientific reports. 3, 1071. 3. Lakaye, B., Makarchikov, A. F., Antunes, A. F., Zorzi, W., Coumans, B., De Pauw, E., Wins, P., Grisar, T. & Bettendorff, L. (2002) J. Biol. Chem. 277, 13771-7. [less ▲]

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See detailCryopreservation of embryos : a way to reduce the number of housed animals and the genetic drift.
Remy, Benoît ULg; Ectors, Fabien ULg; Drion, Pierre ULg

Poster (2014, January 27)

The GIGA Mouse facility platform has recently improved its mouse line cryopreservation technique. The method of embryo cryopreservation by rapid cooling also called aseptic vitrification has been selected ... [more ▼]

The GIGA Mouse facility platform has recently improved its mouse line cryopreservation technique. The method of embryo cryopreservation by rapid cooling also called aseptic vitrification has been selected. Vitrification media, key steps and timing have been optimized and validated. After a first partial exposition of the embryos to cryoprotective solutions, they are immersed in a vitrifying mixture of penetrating and non-penetrating cryoprotectants for a short time. The straw containing the embryos is immediately sealed before to be plunged in LN2, resulting in a brutal solidification in which crystallization does not have time to occur. [less ▲]

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See detailGIGA ANIMAL CARE : Mice & Zebrafish Animal Facility and Transgenesis
Remy, Benoît ULg; Ectors, Fabien ULg; Winandy, Marie ULg et al

Poster (2014, January 27)

In fundamental research, animal models allow to place molecular and cellular observations back into their physiological context. In applied research, these models still remain a mandatory step to evaluate ... [more ▼]

In fundamental research, animal models allow to place molecular and cellular observations back into their physiological context. In applied research, these models still remain a mandatory step to evaluate the efficiency and the toxicity of potential treatments, before going to clinical trials. Mouse and Zebrafish (Danio rerio) are two very interesting models because of a short live cycle and a high prolificacy. They require a limited space. Their genome is well known and shows a high homology with the human. Many tools are available to produce transgenic mice or zebrafishes. Many tests are validated using both these species. [less ▲]

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See detailVitrification of immature equine oocytes
Gatez, Carine ULg; Ectors, Fabien ULg; Farnir, Frédéric ULg et al

Poster (2013, October 11)

Detailed reference viewed: 37 (6 ULg)