Abstract
Hepatocytes from adult rats were maintained in primary culture for up to 10-13 days on nylon meshes coated with a thin layer of rat tail collagen gel. Their ultrastructure closely resembled that of the liver parenchymal cell in vivo, but hepatocytes in late culture exhibited a pronounced buildup of microfilaments beneath their apical cell surface. Hepatocytes in early and late cultures secreted albumin, transferrin, and alpha1-acid glycoprotein into the medium; they exhibited a 7- to 10-fold induction of tyrosine aminotransferase activity by dexamethasone; and they expressed an alkaline phosphatase that was similar to that of normal rat liver with respect to its inhibition by the liver enzyme inhibitor L-homoarginine. In addition, the hepatocytes in culture demonstrated phenotypic changes characteristic of fetal liver parenchymal cells. These changes, which paralleled an increase in DNA synthesis, included the expression of and linear increase in the activity of the fetal liver cell enzyme gamma-glutamyl transpeptidase, an increased production of alpha1-fetoprotein, and a change in the substrate specificity of fructose-bisphosphate aldolase to that of the fetal liver isozyme.
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- BOETTCHER E. W., KISTLER P., NITSCHMANN H. Method of isolating the beta1-metal-combining globulin from human blood plasma. Nature. 1958 Feb 15;181(4607):490–491. doi: 10.1038/181490b0. [DOI] [PubMed] [Google Scholar]
- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bernaert D., Wanson J. C., Drochmans P., Popowski A. Effect of insulin on ultrastructure and glycogenesis in primary cultures of adult rat hepatocytes. J Cell Biol. 1977 Sep;74(3):878–900. doi: 10.1083/jcb.74.3.878. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bissell D. M., Hammaker L. E., Meyer U. A. Parenchymal cells from adult rat liver in nonproliferating monolayer culture. I. Functional studies. J Cell Biol. 1973 Dec;59(3):722–734. doi: 10.1083/jcb.59.3.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cereijido M., Robbins E. S., Dolan W. J., Rotunno C. A., Sabatini D. D. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. doi: 10.1083/jcb.77.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chapman G. S., Jones A. L., Meyer U. A., Bissell D. M. Parenchymal cells from adult rat liver in nonproliferating monolayer culture. II. Ultrastructural studies. J Cell Biol. 1973 Dec;59(3):735–747. doi: 10.1083/jcb.59.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chedid A., Nair V. Ontogenesis of cytoplasmic organelles in rat hepatocytes and the effects of prenatal phenobarbital on endoplasmic reticulum development. Dev Biol. 1974 Jul;39(1):49–62. doi: 10.1016/s0012-1606(74)80007-2. [DOI] [PubMed] [Google Scholar]
- Cheng S., Nassar K., Levy D. gamma-Glutamyltranspeptidase activity in normal, regenerating and malignant hepatocytes. FEBS Lett. 1978 Jan 15;85(2):310–312. doi: 10.1016/0014-5793(78)80480-3. [DOI] [PubMed] [Google Scholar]
- Fiala S., Mohindru A., Kettering W. G., Fiala A. E., Morris H. P. Glutathione and gamma glutamyl transpeptidase in rat liver during chemical carcinogenesis. J Natl Cancer Inst. 1976 Sep;57(3):591–598. doi: 10.1093/jnci/57.3.591. [DOI] [PubMed] [Google Scholar]
- KAMIYAMA S., SCHMID K. Studies on the structure of alpha1-acid glycoprotein. II. Preparation and characterization of a glycopeptide fraction. Biochim Biophys Acta. 1962 Mar 26;58:80–87. doi: 10.1016/0006-3002(62)90819-3. [DOI] [PubMed] [Google Scholar]
- KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
- Kletzien R. F., Pariza M. W., Becker J. E., Potter V. R., Butcher F. R. Induction of amino acid transport in primary cultures of adult rat liver parenchymal cells by insulin. J Biol Chem. 1976 May 25;251(10):3014–3020. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lebherz H. G., Rutter W. J. The class I (Schiff base) fructose-diphosphate aldolase of Peptococcus aerogenes. Methods Enzymol. 1975;42:249–258. doi: 10.1016/0076-6879(75)42122-x. [DOI] [PubMed] [Google Scholar]
- Leffert H., Moran T., Sell S., Skelly H., Ibsen K., Mueller M., Arias I. Growth state-dependent phenotypes of adult hepatocytes in primary monolayer culture. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1834–1838. doi: 10.1073/pnas.75.4.1834. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
- Michalopoulos G., Pitot H. C. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp Cell Res. 1975 Aug;94(1):70–78. doi: 10.1016/0014-4827(75)90532-7. [DOI] [PubMed] [Google Scholar]
- Rutenburg A. M., Kim H., Fischbein J. W., Hanker J. S., Wasserkrug H. L., Seligman A. M. Histochemical and ultrastructural demonstration of gamma-glutamyl transpeptidase activity. J Histochem Cytochem. 1969 Aug;17(8):517–526. doi: 10.1177/17.8.517. [DOI] [PubMed] [Google Scholar]
- Santerre R. F., Rich A. Actin accumulation in developing chick brain and other tissues. Dev Biol. 1976 Nov;54(1):1–12. doi: 10.1016/0012-1606(76)90281-5. [DOI] [PubMed] [Google Scholar]
- Sattler C. A., Michalopoulos G., Sattler G. L., Pitot H. C. Ultrastructure of adult rat hepatocytes cultured on floating collagen membranes. Cancer Res. 1978 Jun;38(6):1539–1549. [PubMed] [Google Scholar]
- Savage C. R., Jr, Bonney R. J. Extended expression of differentiated function in primary cultures of adult liver parenchymal cells maintained on nitrocellulose filters. I. Induction of phosphoenolpyruvate carboxykinase and tyrosine aminotransferase. Exp Cell Res. 1978 Jul;114(2):307–315. doi: 10.1016/0014-4827(78)90488-3. [DOI] [PubMed] [Google Scholar]
- Schapira F., Reuber M. D., Hatzfeld A. Resurgence of two fetal-type of aldolases (A and C) in some fast-growing hepatomas. Biochem Biophys Res Commun. 1970 Jul 27;40(2):321–327. doi: 10.1016/0006-291x(70)91012-0. [DOI] [PubMed] [Google Scholar]
- Schreiber G., Rotermund H. M., Maeno H., Weigand K., Lesch R. The proption of the incorporation of leucine into albumin to that into totaprotein in rat liver and hepatoma Morris 5123 TC. Eur J Biochem. 1969 Sep;10(2):355–361. doi: 10.1111/j.1432-1033.1969.tb00698.x. [DOI] [PubMed] [Google Scholar]
- Sell S., Becker F. F., Leffert H. L., Watabe L. Expression of an oncodevelopmental gene product (alpha-fetoprotein) during fetal development and adult oncogenesis. Cancer Res. 1976 Nov;36(11 Pt 2):4239–4249. [PubMed] [Google Scholar]
- Shaw J. L., Blanco J., Mueller G. C. Simple procedure for isolation of DNA, RNA and protein fractions from cultured animal cells. Anal Biochem. 1975 May 12;65(1-2):125–131. doi: 10.1016/0003-2697(75)90498-4. [DOI] [PubMed] [Google Scholar]
- Sirica A. E., Goldblatt P. J., McKelvy J. F. Isolation and partial characterization of a bile canalicular plasma membrane fraction from normal and regenerating rat liver. J Biol Chem. 1975 Aug 25;250(16):6464–6468. [PubMed] [Google Scholar]
- Tateishi N., Higashi T., Nomura T., Naruse A., Nakashima K. Higher transpeptidation activity and broad acceptor specificity of gamma-glutamyltransferases of tumors. Gan. 1976 Apr;67(2):215–222. [PubMed] [Google Scholar]
- Walker P. R., Potter V. R. Isozyme studies on adult, regenerating, precancerous and developing liver in relation to findings in hepatomas. Adv Enzyme Regul. 1972;10:339–364. doi: 10.1016/0065-2571(72)90022-2. [DOI] [PubMed] [Google Scholar]
- Watabe H. Purification and chemical characterization of alpha-fetoprotein from rat and mouse. Int J Cancer. 1974 Mar 15;13(3):377–388. doi: 10.1002/ijc.2910130313. [DOI] [PubMed] [Google Scholar]


