title: Generation of cloned and chimeric embryos/offspring using the new methods of animal biotechnology.
abstract: The article summarizes results of studies concerning: 1/ qualitative evaluation of pig nuclear donor cells to somatic cell cloning, 2/ developmental potency of sheep somatic cells to create chimera, 3/ efficient production of chicken chimera. The quality of nuclear donor cells is one of the most important factors to determine the efficiency of somatic cell cloning. Morphological criteria commonly used for qualitative evaluation of somatic cells may be insufficient for practical application in the cloning. Therefore, different types of somatic cells being the source of genomic DNA in the cloning procedure were analyzed on apoptosis with the use of live-DNA or plasma membrane fluorescent markers. It has been found that morphological criteria are a sufficient selection factor for qualitative evaluation of nuclear donor cells to somatic cell cloning. Developmental potencies of sheep somatic cells in embryos and chimeric animals were studied using blastocyst complementation test. Fetal fibroblasts stained with vital fluorescent dye and microsurgically placed in morulae or blastocysts were later identified in embryos cultured in vitro. Transfer of Polish merino blastocysts harbouring Heatherhead fibroblasts to recipient ewes brought about normal births at term. Newly-born animals were of merino appearance with dark patches on their noses, near the mouth and on their clovens. This overt chimerism shows that fetal fibroblasts introduced to sheep morulae/blastocysts revealed full developmental plasticity. To achieve the efficient production of chicken chimeras, the blastodermal cells from embryos of the donor breeds, (Green-legged Partridgelike breed or GPxAraucana) were transferred into the embryos of the recipient breed (White Leghorn), and the effect of chimerism on the selected reproductive and physiological traits of recipients was examined. Using the model which allowed identification of the chimerism at many loci, it has been found that 93.9% of the examined birds were chimeras. The effect of donor cells on the reproduction and physiology of the recipients was evident.
authors.affiliation: Department of Animal Reproduction Biotechnology, National Research Institute of Animal Production, Balice/Kraków, Poland. mskrzysz@izoo.krakow.pl
authors.forename: Maria
authors.initials: M
authors.lastname: Skrzyszowska
authors.forename: Jolanta
authors.initials: J
authors.lastname: Karasiewicz
authors.forename: Marek
authors.initials: M
authors.lastname: Bednarczyk
authors.forename: Marcin
authors.initials: M
authors.lastname: Samiec
authors.forename: Zdzisław
authors.initials: Z
authors.lastname: Smorag
authors.forename: Bogusław
authors.initials: B
authors.lastname: Waś
authors.forename: Andrzej
authors.initials: A
authors.lastname: Guszkiewicz
authors.forename: Maciej
authors.initials: M
authors.lastname: Korwin-Kossakowski
authors.forename: Maria
authors.initials: M
authors.lastname: Górniewska
authors.forename: Ewa
authors.initials: E
authors.lastname: Szablisty
authors.forename: Jacek A
authors.initials: JA
authors.lastname: Modliński
authors.forename: Paweł
authors.initials: P
authors.lastname: Łakota
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wawrzyńska
authors.forename: Andrzej
authors.initials: A
authors.lastname: Sechman
authors.forename: Dorota
authors.initials: D
authors.lastname: Wojtysiak
authors.forename: Anna
authors.initials: A
authors.lastname: Hrabia
authors.forename: Maria
authors.initials: M
authors.lastname: Mika
authors.forename: Mirosław
authors.initials: M
authors.lastname: Lisowski
authors.forename: Przemysław
authors.initials: P
authors.lastname: Czekalski
authors.forename: Janusz
authors.initials: J
authors.lastname: Rzasa
authors.forename: Ewa
authors.initials: E
authors.lastname: Kapkowska
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002478:Cells, Cultured; D002678:Chimera / Q000502:physiology; D019976:Cloning, Organism / Q000379:methods*