Analysis of plasma protein and lipoprotein synthesis in long-term primary cultures of baboon hepatocytes maintained in serum-free medium
| Autor: | G. Con Smith, Larry E. Estlack, Robert E. Lanford, K. D. Carey, Rick V. Hay |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
Very low-density lipoprotein
Time Factors Lipoproteins Clinical Biochemistry Immunoblotting Plant Science Biology Lipoproteins VLDL chemistry.chemical_compound Tissue culture High-density lipoprotein medicine Centrifugation Density Gradient Animals Secretion Cells Cultured Cell Biology Blood Proteins Blood proteins Molecular biology Culture Media Lipoproteins LDL medicine.anatomical_structure chemistry Biochemistry Liver Low-density lipoprotein Hepatocyte lipids (amino acids peptides and proteins) Lipoproteins HDL Cell Division Biotechnology Developmental Biology Lipoprotein Papio |
| Zdroj: | In vitro cellulardevelopmental biology : journal of the Tissue Culture Association. 25(2) |
| ISSN: | 0883-8364 |
| Popis: | The analysis of lipoprotein synthesis and secretion in primary hepatocytes has been restricted by the short-term viability and low proliferative response of hepatocytes in vitro. During this investigation a serum-free medium formulation was developed that supports long-term maintenance (greater than 70 d) and active proliferation of primary baboon hepatocytes. Examination of proliferating cells by electron microscopy revealed a distinctive hepatocyte ultrastructure including intercellular bile canaliculi and numerous surface microvilli. High levels of secreted apolipoproteins A-I and E were detected in the tissue culture medium by gel electrophoresis and immunoblot analysis. Immunoprecipitation of proteins from [35S]-methionine labeled tissue culture medium revealed the synthesis and secretion of numerous plasma proteins. Metabolic labeling of cells with [35S]-methionine followed by single-spin density gradient flotation of the media demonstrated that apolipoproteins were being secreted in the form of lipoprotein particles with buoyant densities corresponding to the very low density lipoprotein and low density lipoprotein range, and to the high density lipoprotein range. The labeled apolipoproteins included Bh, E, and A-I. This system for primary hepatocyte culture should prove very useful in future investigations on the regulation of lipoprotein production by hepatocytes. |
| Databáze: | OpenAIRE |
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