Development of bovine embryos on Vero/BRL cell monolayers (mixed co-culture)

Duszewska, A. M.; Reklewski, Z.; Pieńkowski, Marek; Karasiewicz, Jolanta; Modliński, Jacek A.

doi:10.1016/s0093-691x(00)00430-1

Cited by 25 publications

(16 citation statements)

References 34 publications

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“…After 168 h of culture, a higher percentage of morulae and blastocysts were observed in the control (39.06%) than in injected group (23.23%). Corresponding results for our previous studies were 44 and 32.2% (Duszewska et al, 2000;Rosochacki et al, 2001). The greater negative effect on embryo development in the present study may be due to the increase in microinjected DNA concentration (3 ng in our previous study vs 5 ng in this one), an observation that was also shown by Behboodi et al (2001).…”

Section: The Influence Of Gfp Construct Injection On the Development supporting

confidence: 88%

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The use of green ﬂuorescent protein (GFP) to select bovine embryos

Duszewska¹,

Козикова²,

Szydlik³

et al. 2003

J. Anim. Feed Sci.

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A major factor limiting the transgenesis in domestic animals is the inefficiency of maintaining large numbers of recipients carrying nontransgenic foetuses. The objectives of this study were: 1. to determine the influence of green fluorescent protein (GFP) construct injection on the development of bovine embryos, 2. to identify and select the GFP positive bovine embryos, and 3. to determine the rate of mosaicism in transgenic embryos. Cattle oocytes were matured and fertilised in vitro and zygotes were microinjected with pCX-EGFP construct consisting of CMV-IE enhancer, chicken β-actin promoter, cDNA of GFP (EGFP-732 bp) and rabbit β-globin polyadenylation sequences. Embryos from control (64) and microinjected (198) groups were cultured in vitro. After 168 h of culture, morula and blastocysts were observed in 39.06% of control and in 23.23% of injected group. We obtained three GFP positive embryos (1.51% of injected zygotes and 6.52% of morulae/blastocysts). One of them was 100, second 75 and third 25% GFP positive (66.7% of mosaicism). Use of gfp gene reporter to select bovine embryos is useful method to increase transgenic offspring, because GFP marker allows to choice only transgenic embryos and transfer them to recipients.

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Section: The Influence Of Gfp Construct Injection On the Development supporting

confidence: 88%

“…The culture medium was renewed twice during this time. After 48 h of culture, 20 µl of medium was removed and 20 µl of fresh Menezo B-2 medium supplemented with 10% FBS was added but after 144 h of culture, 20 µl of medium was removed and 20 µl of Menezo B 2 was added (Duszewska et al, 2000).…”

Section: In Vitro Culture Of Bovine Embryosmentioning

confidence: 99%

The use of green ﬂuorescent protein (GFP) to select bovine embryos

Duszewska¹,

Козикова²,

Szydlik³

et al. 2003

J. Anim. Feed Sci.

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“…After manipulation the embryos were placed on Vero/BRL monolayer (Duszewska et al, 2000) or STO feeder layer to be cultured overnight (for transfer) or longer. The culture medium was DMEM, which was supplemented like for embryonic cells.…”

Section: Co-culture In Vitromentioning

confidence: 99%

Experimental embryonic-somatic chimaerism in the sheep confirmed by random amplified polymorphic DNA assay

Karasiewicz¹,

Sacharczuk²,

Wąs³

et al. 2008

Int. J. Dev. Biol.

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Developmental potencies of sheep somatic cells (foetal fibroblasts, FFs) in chimaeric animals were analysed. FFs from pigmented Polish Heatherhead (wrzosowka) breed were microsurgically injected into morulae or blastocysts of white Polish Merino breed (5 cells to each embryo). In one experiment the cells were stained with vital fluorescent dye PKH26, and chimaeric blastocysts were cultured in vitro to confirm the presence of fluorescent cells. In the majority of experiments the blastocysts were transferred to synchronized recipient ewes for development until term. Cultured embryonic cells (CEC), earlier known to produce chimaeras, were injected into blastocysts in control experiments. Seven young were born from FF-injected embryos and three were born from CEC-injected ones. All of them were white, but all three control lambs and three experimental lambs showed small areas of skin pigmentation, which indicated Heatherhead CEC or FF contribution. Tissue samples originating from three germ layers were taken from two FFsoriginating presumably chimaeric lambs (male and female) at the age of one month for DNA analysis. The random amplified polymorphic DNA-PCR method supplied two markers of chimaerism, which were amplification products of 643 bp and 615 bp long DNA fragments, found in tissues of experimental lambs as well as in FFs, but not in the blood of parents of blastocysts. The 643 bp marker was found in the majority of tissues of both lambs. The 615 bp amplicon was detected in the skin and lungs of the female lamb and in the hooves of the male lamb. Our data show that foetal fibroblasts introduced to sheep blastocysts can participate in development and can contribute to all tissue lineages up to at least one month of age.

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“…In vitro production of bovine embryos (from slaughterhouse oocytes or those recovered by OPU -ovum pick up) under various culture conditions and in co-culture with various types of cells (Duszewska et al, 2000).…”

Section: Applicationsmentioning

confidence: 99%

Experimental embryology of mammals at the Jastrzebiec Institute of Genetics and Animal Breeding

Karasiewicz¹,

Andrzej-Modlinski²

2008

Int. J. Dev. Biol.

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Our Department of Experimental Embryology originated from The Laboratory ofEmbryo Biotechnology, which was organized and directed by Dr. Maria Czlonkowska until her premature death in 1991. Proving successful international transfer of frozen equine embryos and generation of an embryonic sheep-goat chimaera surviving ten years were outstanding achievements of her term. In the 1990s, we produced advanced fetuses of mice after reconstructing enucleated oocytes with embryonic stem (ES) cells, as well as mice originating entirely from ES cells by substitution of the inner cell mass with ES cells. Attempts at obtaining ES cells in sheep resulted in the establishment of embryo-derived epithelioid cell lines from Polish Heatherhead and Polish Merino breeds, producing overt chimaeras upon blastocyst injection. Successful recloning was achieved from 8-cell rabbit embryos, and healthy animals were born from the third generation of cloned embryos. Recently mice were born after transfer of 8-cell embryonic nuclei into selectively enucleated zygotes, and mouse blastocysts were produced from selectively enucleated germinal vesicle oocytes surrounded by follicular cells, upon their reconstruction with 2-cell nuclei and subsequent activation. Embryonic-somatic chimaeras were born after transfer of foetal fibroblasts into 8-cell embryos (mouse) and into morulae and blastocysts (sheep). We also regularly perform the following applications: in vitro production of bovine embryos from slaughterhouse oocytes or those recovered by ovum pick up; cryopreservation of oocytes and embryos (freezing: mouse, rabbit, sheep, goat; vitrification: rabbit, cow); and banking of somatic cells from endangered wild mammalian species (mainly Cervidae).

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Development of bovine embryos on Vero/BRL cell monolayers (mixed co-culture)

Cited by 25 publications

References 34 publications

The use of green ﬂuorescent protein (GFP) to select bovine embryos

The use of green ﬂuorescent protein (GFP) to select bovine embryos

Experimental embryonic-somatic chimaerism in the sheep confirmed by random amplified polymorphic DNA assay

Experimental embryology of mammals at the Jastrzebiec Institute of Genetics and Animal Breeding

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