Expression of insulin-like growth factor IGF-I and IGF-binding proteins in primary porcine hepatocyte culture

Kim I. A., . and Jin E. J., . and Lee C. Y., . (2004) Expression of insulin-like growth factor IGF-I and IGF-binding proteins in primary porcine hepatocyte culture. [Proceedings Paper]

It is known in the rat but not in any other mammalian species that circulating insulin-like growth factors IGFs and multiple species of IGF-binding proteins IIGFBPs are secreted from hepatocytes whereas the major plasma IGF carrier IGFBP-3 is secreted only from non-parenchymal cells in the liver. The objective of the present study was to characterize the expression of IGF-I and IGFBPs in primary porcine hepatocyte culture. Hepatocytes were prepared from a weanling piglet by the collagenase perfusion method. Upon Western ligand blotting using 125IIGF-II the hepatocyte conditioned culture medium CCM was found to contain all the different sizes of IGFBPs found in serum including a 43- and 39-kDa doublet 34-kDa a 31kDa doublet and a 26-kDa IGFBPs. The amounts of these IGFBPs in CCM did not increase after 24-h culture up to 72 h. Moreover the 43-and 39-kDa doublet and 34-kDa IGFBPs were identified as IGFBP-3 and IGFBP-2 respectively by immunoprecipitation with corresponding antisera followed by Western ligand blotting. In line with this the expression of IGFBP-3 and IGFBP-2 as well as IGF-I in cultured hepatocytes was identified by reverse transcription-polymerase chain reaction. These results suggest but do not prove that unlike the rat hepatocyte porcine hepatocyte may express and secrete IGFBP-3.

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Abstract

It is known in the rat but not in any other mammalian species that circulating insulin-like growth factors IGFs and multiple species of IGF-binding proteins IIGFBPs are secreted from hepatocytes whereas the major plasma IGF carrier IGFBP-3 is secreted only from non-parenchymal cells in the liver. The objective of the present study was to characterize the expression of IGF-I and IGFBPs in primary porcine hepatocyte culture. Hepatocytes were prepared from a weanling piglet by the collagenase perfusion method. Upon Western ligand blotting using 125IIGF-II the hepatocyte conditioned culture medium CCM was found to contain all the different sizes of IGFBPs found in serum including a 43- and 39-kDa doublet 34-kDa a 31kDa doublet and a 26-kDa IGFBPs. The amounts of these IGFBPs in CCM did not increase after 24-h culture up to 72 h. Moreover the 43-and 39-kDa doublet and 34-kDa IGFBPs were identified as IGFBP-3 and IGFBP-2 respectively by immunoprecipitation with corresponding antisera followed by Western ligand blotting. In line with this the expression of IGFBP-3 and IGFBP-2 as well as IGF-I in cultured hepatocytes was identified by reverse transcription-polymerase chain reaction. These results suggest but do not prove that unlike the rat hepatocyte porcine hepatocyte may express and secrete IGFBP-3.

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Item Type:	Proceedings Paper
Additional Information:	Summary En
AGROVOC Term:	INSULIN-LIKE GROWTH FACTOR
AGROVOC Term:	PROTEIN CONTENT
AGROVOC Term:	SWINE
AGROVOC Term:	PIGLETS
AGROVOC Term:	PROTEASES
AGROVOC Term:	LIGANDS
AGROVOC Term:	GROWTH FACTORS
Geographical Term:	MALAYSIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:27
URI:	http://webagris.upm.edu.my/id/eprint/16366

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