• Title/Summary/Keyword: hepatocyte primary culture

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Organ Induction by Combined Dose of bFGF and HGF in Animal Cap Assay of Early Xenopus laevis Embryos. (Xenopus laevis 초기 배의 동물극 분리배양에서 bFGF와 HGF 혼합처리에 의한 기관유도)

  • 진정효;윤춘식;이호선;박용욱;정선우
    • Journal of Life Science
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    • v.14 no.3
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    • pp.375-384
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    • 2004
  • Fibroblast growth factors (FGFs) are known to induce multiple functions in early development, including mesoderm formation, gastrulation movement and antero-posterior patterning. The induction of mesoderm from Xenopus presumptive ectoderm and the combination effect on inducing organs of bFGF(basic FGF) and HGF (Hepatocyte Growth Factor) were studied. Explants were cultured in the combined solution for 3 days to normal embryo arrive at St. 43. These effects on combined dose were examined by histological experiment and by immunohistochemical method. The concentrations of growth factors were tested in 0, 0.5, 1, 10 and also tested in 50 ng/ml of bFGF, and 0, 1, 10, 50 and 100ng/ml of HGF respectively. The synergistic effects were seen in the combined-dose of bFGF and HGF rather than in single dose. Various organs were differentiated and highest inducing effects were seen at the combined concentration of 1 ng/ml of bFGF and 10ng/ml of HGF, and at the concentration 10ng/ml of bFGF and 1 ng/ml of HGF. The bFGF induces various organs from cultured animal cap explants and the effects are time and dose-dependent. HGF is also a potent mitogen for renal tubular cells and for mature hepatocytes in primary culture. Eyes were developed in high percentage at the combined concentration of 1 and 10ng/ml of bFGF, and 1 and 10 ng/ml of HGF. From the induced eye and normal embryonic eye, RPE65 was commonly detected by monoclonal antibodies 40All and 25F5 and the localization of RPE65 was seen by AP reaction.

Effect of 2,4-Dichlorophenoxy Acetic Acid on Vitellogenin Synthesis and $E_2$-ER Binding Affinity of Hepatocytes in Rainbow Trout (Oncorhynchus mykiss) (무지개송어, Oncorhynchus mykiss 배양 간세포에서 2,4-Dichlorophenoxy acetic acid이 Vitellogenin 합성과 $E_2$-ER Binding Affinity에 미치는 영향)

  • 황운기
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.31-37
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    • 2002
  • Effect of 2,4-Dichlorophenoxy acetic acid (2,4-D) on vitellogenin (VTG) production and estrogen ($E_2$)-estrogen receptor (ER) binding affinity were examined in primary hepatocyte cultures of rainbow trout, Oncorhynchus mykiss. Hepatocytes were pre-cultured for 2 days; subsequently, $E_2$( 2$\times$$10^{-6}$/ M) and 2,4-D ($10^{-9}~10^{-6}/M$) were simultaneously added to the incubation medium. They were cultured for more than 5 days. VTG and $E_2$-ER binding affinities were analyzed by SDS-PAGE and ELISA, respectively. 2,4-D concentration used had no appreciable effect on the morphology, viability, and DNA content of hepatocytes in culture. It had also no effect on VTG production. However, it interfered with $E_2$-ER binding affinity, which was reduced with increasing concentration of 2,4-D. The affinity was inhibited by 25 and 30% at $10^{-7}$ M and $10^{-6}$ M of 2,4-D, respectively. This result suggested that although 2,4-D had no effect on VTG production, it acted as reno-estrogenic contaminant in ER.