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Asian-Australas J Anim Sci > Volume 21(9); 2008 > Article
Animal Reproduction and Physiology
Asian-Australasian Journal of Animal Sciences 2008;21(9): 1244-1251.
DOI: https://doi.org/10.5713/ajas.2008.70558    Published online September 3, 2008.
Expression and Secretion of the Insulin-like Growth Factor System Components by Pig Liver Cells
I. Kim, E. J. Jin, K. Baik, C. H. Park, W. K. Kim, C. W. Kang, Y. Ko, I. Jang, W. S. Choi, C. Y. Lee*
Correspondence:  C. Y. Lee,
Abstract
The aim of the present study was to delineate the expression and secretion of insulin-like growth factor (IGF) system components by pig liver cells. Hepatocytes were prepared from 3-wk-old weanling piglets following a two-step collagenase perfusion procedure, after which the cells were incubated for 24 or 48 h at a density of 2105 cells per 35-mm dish in 2-ml Williams’ medium E. The cells were found to express the genes encoding IGF-I, IGF-binding proteins (IGFBPs)-2 and -3 and acid-labile subunit (ALS) by reverse transcription-polymerase chain reaction (RT-PCR) following the culture. However, IGF-I was localized to hepatocytes by immunohistochemical analysis, whereas IGFBP-3 was localized to endothelial cells, but not to hepatocytes. This indicated that the IGFBP-3 gene expression detected by RT-PCR was likely to have been contributed by unidentified non-parenchymal cells that had not been removed during the hepatocyte preparation. The conditioned culture medium (CCM) of the cells contained immunoreactive IGF-I and IGF-II, with the latter being seven-fold more abundant than the former. The CCM also contained 43-, 40-, 34-, 31-kDa doublet and 26-kDa IGFBPs as examined by Western ligand blotting. The 40-, 34- and 31-kDa doublet IGFBPs were approximately three-fold as abundant as the 43- and 26-kDa IGFBPs. Moreover, the 43- and 40-kDa doublet and the 34-kDa IGFBPs were immunoprecipitable with IGFBP-3 and IGFBP-2 antibodies, respectively. Overall, these results are similar to those known in the rat, which suggests that the IGF system components are likely to be expressed and secreted in pig liver in a manner similar to that in rat liver.
Keywords: IGF; IGFBP; Gene Expression; Hepatocyte; Pig


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