Importance of cell aggregation for expression of liver functions and regeneration demonstrated with primary cultured hepatocytes
- 1 September 1993
- journal article
- research article
- Published by Wiley in Journal of Cellular Physiology
- Vol. 156 (3), 522-530
- https://doi.org/10.1002/jcp.1041560311
Abstract
Adult rat hepatocytes aggregated to form floating multicellular spheroids when cultured in Primaria dishes, which have a positively charged surface, in serum‐free Williams' medium E (WE) supplemented with insulin and epidermal growth factor (EGF). These hormones were essential for maintenance of the spheroids, whereas the size of the spheroids depended on the inoculum cell density. The spheroids retained in vivo levels of expressions of albumin and glucokinase and synthesized scarcely any DNA even in the presence of insulin and EGF. On transfer to type I collagen‐coated dishes, the spheroids gradually disaggregated and the cells formed monolayers, in which the expressions of albumin and glucokinase were suppressed and DNA synthesis and hexokinase activity were increased. DNA synthesis of hepatocytes in monolayer culture was maximal 24 hr after transfer of the spheroids, ∼80% of the hepatocyte nuclei were labelled with bromodeoxyuridine during culture for 48 hr, and the mitotic index was ∼70% after 60 hr. These results suggest that, in spheroids, hepatocytes remained in the G0 phase, but that when they formed monolayers, they progressed to the G1 phase and proceeded through the cell cycle in the presence of insulin and EGF. This work shows that the cell cycle of hepatocytes in culture can be manipulated by providing conditions for quiescence as spheroids or growth as monolayers and that the shape of hepatocytes is important for regulating their growth and liver‐specific functions.This publication has 28 references indexed in Scilit:
- Cell-density dependent expression of the c-myc gene in primary cultured rat hepatocytesBiochemical and Biophysical Research Communications, 1991
- Multiple sequential periods of DNA synthesis and quiescence in primary hepatocyte cultures maintained on the DMSO‐EGF on/off protocolJournal of Cellular Physiology, 1989
- Interleukin-1β is a potent growth inhibitor of adult rat hepatocytes in primary cultureExperimental Cell Research, 1988
- Primary hepatocyte culture: Is it home away from home?Hepatology, 1988
- Proteoglycans and glycosaminoglycans induce gap junction synthesis and function in primary liver cultures.The Journal of cell biology, 1987
- In vitro induction of neonatal rat hepatocytes by direct contact with adult rat hepatocytes*1Experimental Cell Research, 1987
- Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver.JCI Insight, 1987
- Glycosaminoglycans and proteoglycans induce gap junction expression and restore transcription of tissue-specific mRNAs in primary liver culturesHepatology, 1987
- Spheroidal aggregate culture of rat liver cells: histotypic reorganization, biomatrix deposition, and maintenance of functional activities.The Journal of cell biology, 1985
- Control of growth and expression of differentiated functions of mature hepatocytes in primary culture.Cell Structure and Function, 1985