Synthesis and Secretion of Lipoproteins by Human Hepatocytes in Culture

• Published in: In Vitro Cellular & Developmental Biology 1988-02, Vol.24 (2), p.85-90
• Main Authors: M. E. Bouma, M. Pessah, Renaud, G., N. Amit, D. Catala, R. Infante
• Format: Article
• Language: English
• Subjects: Animal cells; Apolipoprotein A-I; Apolipoproteins - biosynthesis; Apolipoproteins - secretion; Apolipoproteins A - biosynthesis; Apolipoproteins A - secretion; Apolipoproteins B - biosynthesis; Apolipoproteins B - secretion; Apolipoproteins C - biosynthesis; Apolipoproteins C - secretion; Apolipoproteins E - biosynthesis; Apolipoproteins E - secretion; Apoproteins; Autoradiography; Biological and medical sciences; Biotechnology; Cell cultures. Hybridization. Fusion; Cell Survival; Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; HDL lipoproteins; Hepatocellular carcinoma; Hepatocytes; Histocytochemistry; Humans; Immunoassay; Immunoenzyme Techniques; Lipids; Lipoproteins; Lipoproteins - biosynthesis; Lipoproteins - secretion; Lipoproteins, HDL - biosynthesis; Lipoproteins, HDL - secretion; Lipoproteins, LDL - biosynthesis; Lipoproteins, LDL - secretion; Lipoproteins, VLDL - biosynthesis; Lipoproteins, VLDL - secretion; Liver; Liver - cytology; Liver - metabolism; Liver - secretion; Methods. Procedures. Technologies; Miscellaneous; Models, Biological; Molecular and cellular biology; Monoclonal antibodies; Polyclonal antibodies; Protein Biosynthesis; RNA, Messenger - genetics; Secretion
• ISSN: 0883-8364; 2327-431X; 1475-2689
• DOI: 10.1007/BF02623884

Confluent monolayers of normal human hepatocytes obtained by collagenase perfusion of liver fragments were incubated in a serum-free medium. Intracellular apolipoproteins apo AI, apo C, apo B, and apo E were detected between Day 1 and Day 6 of the culture by immunoenzymatic staining using polyclonal antibodies directed against these apoproteins and monoclonal antibodies directed against both forms of apo B (B100 and B48). Translation of mRNA isolated from these hepatocytes in an acellular system revealed that apo AI and apo E were synthesized as the precusor forms of mature plasma apo AI and apo E. Three lipoprotein fractions corresponding to the density of very low density lipoprotein (VLDL), low density lipoprotein (LDL), and high density lipoprotein (HDL) were isolated from the medium at Day 5 of culture and examined by electron microscopy after negative staining. VLDL and LDL particles are similar in size and shape to plasma lipoproteins; spherical HDL are larger than normal plasma particles isolated at the same density. Their protein represented 44, 19.5, and 36.5% respectively, of the total lipoprotein protein. The secretion rate of VLDL protein corresponded to that measured in primary cultures of rat hepatocytes. After incorporation of$[^3H]glycerol$, more than 92% of the$[^3H]triglyceride$secreted into the medium was recovered in the VLDL fraction. These results demonstrate that primary cultures of normal human hepatocytes are able to synthesize and secrete lipoproteins and thus could be a useful model to study lipoprotein metabolism in human liver.