title: Increased reactivity of cultured chicken blastodermal cells to anti-stage-specific embryonic antigen-1 antibody after exposure to bone morphogenetic proteins.
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 embryo-derived 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 3ng 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.
authors: Department of Food and Animal Biotechnology, Division of Animal Genetic Engineering, Seoul National University, Seoul 151-921, Republic of Korea.
authors: Duk Kyung
authors: DK
authors: Kim
authors: Ki Duk
authors: KD
authors: Song
authors: Jin Nam
authors: JN
authors: Kim
authors: Tae Sub
authors: TS
authors: Park
authors: Jeong Mook
authors: JM
authors: Lim
authors: Jae Yong
authors: JY
authors: Han
mesh_terms: D000818:Animals; D001756:Blastoderm / Q000187:drug effects*; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology*; D004306:Dose-Response Relationship, Immunologic; D016256:Lewis X Antigen / Q000276:immunology*; D010770:Phosphotransferases / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D019485:Bone Morphogenetic Proteins; D016256:Lewis X Antigen; D010770:Phosphotransferases