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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 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 detailLong term culture and characterization of chicken primordial germ cells
Tonus, Céline ULg; Cloquette, Karine; Ectors, Fabien ULg et al

Poster (2013, October)

Avian primordial germ cells (PGCs) can keep their germ cells properties and are foreseen as promising tools for developing avian transgenesis and preservation of genetic resources of endangered species ... [more ▼]

Avian primordial germ cells (PGCs) can keep their germ cells properties and are foreseen as promising tools for developing avian transgenesis and preservation of genetic resources of endangered species. We have developed original methods that allow long term (20 month) expansion of primary cultures of undifferentiated PGCs and their efficient cryopreservation. Blood samples were collected from stage 13-18 embryos, pooled, deposited in cell culture inserts and co-cultivated in the presence of irradiated BRL cells. This physically separated co-culture system along with selective culture medium promoted emergence, selection and proliferation of undifferentiated PGCs lines. Overall, 35% of blood samples gave rise to PGCs cell lines originating from three commercial layer breeds and two Belgian endangered breeds. Moreover, we recently isolate and cultivate a new PGC line from turkey. All PGCs lines were first characterised for the expression of the stem cells and PGCs characteristic 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. In addition, by means of a quantitative PCR amplification of a chromosome W specific sequence, we demonstrated a progressive drift of all our lines towards the male sex (WL), while they were initially isolated from pooled blood samples with statistically equivalent numbers of male and female embryos (35 females: 29 males). PGCs were subsequently efficiently cryopreserved by slow freezing or by a newly developed vitrification method. Labelled PGCs from 10 lines were injected in recipient embryos. At day 6, colonization of the genital ridges confirmed that PGCs retain their gonadal migratory ability, both after long-term culture (min 3, max 20 month) and after cryopreservation. In order to evaluate the germinal differentiation of cultured PGCs during the gonadal development as well as the germline transmission rate, we established a stably expressing GFP line that was successfully injected in emrbyos. Results are in progress. [less ▲]

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See detailLong term culture and characterization of chicken primordial germ cells
Tonus, Céline ULg; Waroux, Olivier ULg; Cloquette, Karine et al

Poster (2012, November)

Avian primordial germ cells (PGCs), can keep their germ cells properties and are foreseen as promising tools for developing avian transgenesis and preservation of genetic resources of endangered species ... [more ▼]

Avian primordial germ cells (PGCs), can keep their germ cells properties and are foreseen as promising tools for developing avian transgenesis and preservation of genetic resources of endangered species. We have developed original methods that allow long term (20 month) expansion of primary cultures of undifferentiated PGCs and their efficient cryopreservation. Blood samples were collected from stage 13-18 embryos, pooled, deposited in cell culture inserts and co-cultivated in the presence of irradiated BRL cells. This physically separated co-culture system along with selective culture medium promoted emergence, selection and proliferation of undifferentiated PGCs lines. Overall, 35% of blood samples gave rise to PGCs cell lines originating from three commercial layer breeds and two Belgian endangered breeds. PGCs lines were first characterised for the expression of the stem cells and PGCs characteristic marker SSEA-1 by FACS (expression rate: 90-99%). RT-PCR confirmed expression of germ-line specific markers (CVH, CDH, DAZL), pluripotency markers (cPouV, cSox2, cNanog), telomerase and CXCR4 receptor. In addition, by means of a quantitative PCR amplification of a chromosome W specific sequence, we demonstrated a drift of all our lines towards the male sex (WL), while they were initially isolated from pooled blood samples with statistically equivalent numbers of male and female embryos (35 females: 29 males). PGCs were subsequently efficiently cryopreserved by slow freezing or by a newly developed vitrification method. Labelled PGCs from 10 lines were injected in recipient embryos. Colonization of the genital ridges confirmed that PGCs retain their gonadal migratory ability, both after long-term culture (min 3, max 20 month) and after cryopreservation. [less ▲]

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See detailPathology of influenza virus H5N1 infection in resistant and susceptible laboratory mice.
Garigliany, Mutien-Marie ULg; Cloquette, Karine; Leroy, Michaël et al

Conference (2008, September)

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See detailLes dynamines MX et leur activité anti-influenza.
Garigliany, Mutien-Marie ULg; Zecchinon, Laurent; Cloquette, Karine et al

Poster (2008, September)

Detailed reference viewed: 7 (2 ULg)
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See detailMX dynamins and the innate resistance opposed to influenza virus.
Garigliany, Mutien-Marie ULg; Baise, Etienne; Cloquette, Karine et al

Conference (2008, March)

Detailed reference viewed: 4 (1 ULg)