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Increased reactivity of cultured chicken blastodermal cells to anti-stage-specific embryonic antigen-1 antibody after exposure to bone morphogenetic proteins

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Authors
Kim, Duk Kyung; Song, Ki Duk; Kim, Jin Nam; Park, Tae Sub; Lim, Jeong Mook; Han, Jae Yong
Issue Date
2006
Publisher
Elsevier
Citation
Theriogenology, vol.65 no.3, pp. 658-668
Keywords
Blastodermal cellStage-specific embryonic antigenBone morphogenetic protein (BMP)BMP receptor kinase (BRK)In vitro-culture
Abstract
We evaluated whether bone morphogenetic proteins (BMPs) increased the reactivity of chicken stage X blastodermal cells to the germ cell marker, anti-stage-specific embryonic antigen (SSEA)-1 antibody. In Experiment 1, blastodermal cells cultured on a feeder layer of SIM mouse embryoderived thioguanine and ouabain resistant (STO) cells were treated with different doses of BMP-2 and/or BMP-4, and the anti-SSEA-1 antibody reactivity of cultured cells was examined 48 h later. A significant (P < 0.05) increase in the number of anti-SSEA-1 antibody-positive cells was detected after the addition of 75 or 100 ng/ml BMP-2. Neither 0–20 ng/ml BMP-4 nor the combined addition of 75 ng/ml BMP-2 with either 10 or 15 ng/ml BMP-4 increased reactivity more than that induced by 75 ng/ml BMP-2 alone. Results of the qualification and quantification of BMP receptor kinase (BRK)-1, BRK-2, and BRK-3 using RT-PCR and real-time PCR showed that all three receptors were detected in blastodermal cells treated with BMPs, intact stage X embryos and 5.5-day-old embryonic gonads, but no expression was detected in STO feeder cells. In Experiment 2, the treatment of stage X embryos with different doses of BMP-2 (0.15–3 ng/embryo) or BMP-4 (0.02–0.4 ng/embryo) did not affect the reactivity of 5.5-day-old embryonic gonadal cells to the anti-SSEA-1 antibody. BRK-1 expression was selectively increased in stage X embryos after the infusion of 3 ng BMP-2 than after no infusion, but no changes in other BRKs’ expression were detected. In conclusion, the addition of BMP-2 to culture medium in the presence of STO feeder cells promoted the reactivity of blastodermal cells to anti-SSEA-1 antibody, which might contribute to the generation of chicken primordial germ cell precursor or germ cell-like cells. The relationship between BMP action and BRK expression was further discussed.
ISSN
0093-691X
Language
English
URI
http://hdl.handle.net/10371/100221
DOI
https://doi.org/10.1016/j.theriogenology.2005.06.008
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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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