Foetal fibroblasts introduced to cleaving mouse embryos contribute to full-term development

Piliszek, Anna; Modliński, Jacek A.; Pyśniak, Kazimiera; Karasiewicz, Jolanta

doi:10.1530/rep-06-0013

Cited by 15 publications

(20 citation statements)

References 71 publications

(51 reference statements)

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“…In few cases the embryos cultured for one week displayed but one marked cell. The observations are not in disagreement with the results of detailed analysis in the mouse (Piliszek et al, 2007), showing that FFs resume divisions at day 3 after injection and that cell loses happen at any stage of development.…”

Section: Presence Of Pkh26-stained Fibroblasts In Blastocystssupporting

confidence: 65%

“…First, experimental reprogramming of cells in vitro culminated in the results by Takahashi and Yamanaka (2006), who have shown that the four genes introduced to foetal or adult fibroblasts caused a small fraction of fibroblasts to become totipotent ES-like cells. The second line of evidence is chimaera formation after injecting foetal fibroblasts to cleaving mouse embryos (Piliszek et al, 2007). Progeny of injected somatic cells could be found in many organs of adult mice, suggesting their reprogramming in vivo and subsequent participation in development.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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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Section: Presence Of Pkh26-stained Fibroblasts In Blastocystssupporting

confidence: 65%

Section: Introductionmentioning

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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“…This means that mechanical damage caused by injection of MBT blastomeres is greater than the damage caused by injection of the same number of blastema cells. The survival at 72 h, in E and the controls (C 1 and C 2 ) ( Piliszek et al, 2007). The results of this work suggest that insertion of blastema cells does not have a negative effect on embryo development for at least three days post fertilization.…”

Section: Resumen Evaluación De Las Células De La Blastema De Aleta Camentioning

confidence: 61%

Short communication. Evaluation of presumptive caudal fin blastema cells as candidate donors in intraspecies zebrafish (Danio rerio) chimaeras

Camps

Simão

Costa

et al. 2008

Span. j. agric. res.

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The blastema is a regenerative tissue with remarkable pluripotency. The aim of this work done on zebrafish (Danio rerio) was to define technical procedures required for obtaining and integrating blastema cells into embryos at the mid blastula transition stage (MBT) and the effect on survival, as well as the capacity to produce pigmented chimaeras. Wild type blastema cells were injected into gold type MBT embryos (E). Wild MBT blastomere cells were also injected into gold type MBT embryos as a control (C 1 ). A second control group, C 2 , was not subjected to any manipulation. Survival was evaluated at 24, 48 and 72 h after performing the chimaerism, and the rate of adult chimaeras evaluated. The results showed significant differences in embryo survival between the E and C 1 groups in embryo survival at 24 and 48 h postchimaerism (24 h: E-83.49% vs C 1 -54.8%, p < 0.05; 48 h: E-98.83% vs C 1 -85.13%, p < 0.05). There was no significant difference, at any time, between E and C 2 . The results at 72 h for E and the controls (E-89.41%; C 1 -84.12% and C 2 -92.55%) indicate that insertion of blastema cells does not have a negative effect on embryo development. The results in adults (E: 0 chimaeras from 7 specimens; C 1 : 5 chimaeras from 17 specimens) suggest that the dedifferentiation grade of the blastema cells may not be enough to generate germ-line chimaeras, but their condition of potentially dedifferentiating cells may be an advantage when using them as donor nuclei in somatic cloning by nuclear transplant.Additional key words: biodiversity, chimaerism, embryo, zebrafish. Resumen Evaluación de las células de la blastema de aleta caudal como donantes en quimeras intraespecíficas de pez cebraEl blastema es un tejido regenerativo con una pluripotencia remarcable. El objetivo del presente trabajo realizado en pez cebra (Danio rerio) es definir los procedimientos técnicos requeridos para la obtención e integración de células de la blastema en embriones en estadio MBT y su efecto en la supervivencia, así como también la capacidad de dar lugar a quimeras pigmentadas. Se inyectaron células de blastema de la estirpe silvestre en embriones gold en estadio MBT (E) y como control se inyectaron blastómeras silvestres MBT en embriones gold MBT (C 1 ). El segundo grupo control (C 2 ), no fue sometido a manipulación. Se evaluó la tasa de supervivencia a 24, 48 y 72 h post-quimerismo y la tasa de quimeras adultas. Los resultados mostraron diferencias significativas en la supervivencia embrionaria a las 24 y 48 h entre el grupo E y el C 1 (24 h: E-83,49% vs C 1 -54,8%, p<0,05; 48 h: E-98,83% vs C 1 -85,13%, p<0,05). No se detectaron diferencias significativas entre los grupos E y C 2 en ningún momento. Los resultados obtenidos a las 72 h, tanto en E como en los controles (E-89,41%; C 1 -84,12% y C 2 -92,55%) sugieren que la inserción de células de blastema no causa efectos negativos en el desarrollo del embrión. Los resultados obtenidos en adultos (E: 0 quimeras de 7 especímenes; C 1 : 5 quimeras de 17 especí-menes) apun...

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“…In the mouse, it was shown by GPI analysis that, starting from the day 11-12 of development, the introduced somatic cells fuse with the host cells, and can persist -as hybrid cells -in tissues derived from all three germ layers (Fig. 8) (Piliszek et al, 2007). Also, in sheep, the presence of progeny of the introduced somatic cells was foundas was shown by RAPD analysis -in several tissues derived from three germ layers (Karasiewicz et al, 2008, pp.…”

Section: Scientific Programmentioning

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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Foetal fibroblasts introduced to cleaving mouse embryos contribute to full-term development

Cited by 15 publications

References 71 publications

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

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

Short communication. Evaluation of presumptive caudal fin blastema cells as candidate donors in intraspecies zebrafish (Danio rerio) chimaeras

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

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