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Establishment of an in vitro culture system for chicken preblastodermal cells

Cited 17 time in Web of Science Cited 16 time in Scopus
Authors
Park, Hyun Jeong; Park, Tae Sub; Kim, Tae Min; Kim, Jin Nam; Shin, Sang Su; Lim, Jeong Mook; Han, Jae Yong
Issue Date
2006
Publisher
Wiley
Citation
Molecular Reproduction and Development, vol.73 no.4, pp. 452-461
Keywords
chimerain vitro culturecolony formationstem cellsubculture
Abstract
To develop an alternative source for chicken pluripotent cells, we examined (1) whether undifferentiated preblastodermal cells could be subcultured in vitro for an extended period and (2) how subculturing affected the physiological properties of preblastodermal cells. The average number of preblastodermal cells was 2,397 in stage V embryos and 36,345 in stage VII embryos; stage X embryos had an average of 53,857 blastodermal cells. The average cell size decreased significantly (70.63–18.83 mm in diameter; P<0.0001) as the embryo grew; this was closely related to a reduction in the size and number of lipid vesicles in the cell cytoplasm. The culture conditions were optimized for the stage V preblastodermal cells and the control stage X blastodermal cells. On STO feeder cells, the preblastodermal cells achieved stable growth in vitro only in HES medium or a mixed medium of the Knockout DMEM and HES media. However, more than 10 passages of preblastodermal cells at intervals of 3–4 days was possible only by using the Knockout/HES mixed medium and BRL cell-conditioned HES medium for the primary cultures and subcultures, respectively. Colony-forming preblastodermal cells had well-delineated cytoplasm, which was positively stained for stem cell-specific markers by anti-stage-specific embryo antigen-1 antibody, periodic acid-Schiffs solution, and alkaline
phosphatase. When preblastodermal cells with or without culturing were transferred into the blastodermal cavity of stage X embryos, only in vitro-cultured preblastodermal cells at stage V (4/5¼80%) and stage VII (2/8¼25%) induced somatic chimerism in recipient chickens. In conclusion, undifferentiated preblastodermal cells could be subcultured, and only the colony-forming preblastodermal cells that stained positively for stem cell markers could induce somatic chimerism.
ISSN
1040-452X
Language
English
URI
https://hdl.handle.net/10371/100222
DOI
https://doi.org/10.1002/mrd.20441
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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