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Agricultural Biology  / №6 2012

ISOLATION AND CHARACTERISTICS OF PRIMORDIAL GERM CELLS IN CHICKEN (150,00 руб.)

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Первый авторTomgorova
АвторыVolkova N.A., Volkova L.A., Fisinin V.I., Zinovieva N.A.
Страниц3
ID436669
АннотацияOptimized conditions for isolation of primordial germ cells in chicken and their description are given. Trypsinization (with 0.05 % trypsin) was found to be the most effective method of disaggregation in chick embryos for isolation of their embryonic cells. The maximum number of primordial germs cells can be isolated from 6 days-old embryos, followed by separation of the resulting cell suspension by adhesion after 90 minutes of their cultivation with the use of chicken embryonic fibroblasts as feeder layer. During incubation of primordial germ cells on feeder layer cells remained viable for a longer time, and ranged from 5 to 7 days, depending on the type of cells used as feeder layer (finite cell line STO, recently separated and incubated for several passages in chicken embryo fibroblasts).
УДК636.52/.58:591.324:576.3/.7.086.83:591.04
ISOLATION AND CHARACTERISTICS OF PRIMORDIAL GERM CELLS IN CHICKEN / Е.К. Tomgorova [и др.] // Agricultural Biology .— 2012 .— №6 .— С. 17-19 .— URL: https://rucont.ru/efd/436669 (дата обращения: 27.04.2024)

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Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2012, № 6, p. 61-65 UDK 636.52/.58:591.324:576.3/.7.086.83:591.04 1 All-Russia Research and Development Institute for Livestock Husbandry, RAAS, Dubrovitsy settlement 142132, Moscow province, Russia e-mail: natavolkova@inbox.ru, n_zinovieva@mail.ru ISOLATION AND CHARACTERISTICS OF PRIMORDIAL GERM CELLS IN CHICKEN Е. <...> Trypsinization (with 0.05 % trypsin) was found to be the most effective method of disaggregation in chick embryos for isolation of their embryonic cells. <...> The maximum number of primordial germs cells can be isolated from 6 days-old embryos, followed by separation of the resulting cell suspension by adhesion after 90 minutes of their cultivation with the use of chicken embryonic fibroblasts as feeder layer. <...> During incubation of primordial germ cells on feeder layer cells remained viable for a longer time, and ranged from 5 to 7 days, depending on the type of cells used as feeder layer (finite cell line STO, recently separated and incubated for several passages in chicken embryo fibroblasts). <...> A significant current advance in creation of transgenic bird has been obtained through the use of various methods of gene transfer (1-7), but production of transgenic chickens is still concerned to certain problems. <...> Peculiarities of bird reproduction and development (8) reduce the efficiency of microinjection commonly performed on animal cells, which necessitates applying some alternative methods of directed gene transfer. <...> One of such methodological approaches is actively developed in recent years – using pluripotent stem cells (PSC) of different types (including primordial germ cells - predecessors of differentiated germ cells) as target cells for the transfer of recombinant DNA (9). <...> Genetic modification of primordial germ cells (PGCs) with following transplantation into chicken embryos in vivo is considered as a promising method for target genetic modification of bird gonads and creation of transgenic bird. <...> In this connection, the subject of this research was isolation and characterization of primordial germ cells of chickens as target cells for genetic engineering. <...> Experiments were performed on chicken embryos the cross Ptichnoe at developmental stages from the 4th to 8th day of incubation. <...> Disaggregation of the embryos was performed using two methods: mechanical dissociation of cells and enzymatic treatment of tissues. <...> Dissociation of the cells was performed <...>