Maintenance of steroid metabolism in primary cultures of adult rat hepatocytes in serum-free medium

1986 ◽  
Vol 14 (5) ◽  
pp. 914-915 ◽  
Author(s):  
ABAS H. HUSSIN ◽  
PAUL SKETT
Keyword(s):  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Steroid Metabolism ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Adult Rat ◽  
Adult Rat Hepatocytes

Improved maintenance of adult rat hepatocytes in a new serum-free medium in the presence or absence of barbiturates

1989 ◽  
Vol 25 (9) ◽  
pp. 839-848 ◽  
Author(s):  
Masahiro Miyazaki ◽  
Yasunori Suzuki ◽  
Munehiro Oda ◽  
Akira Kawai ◽  
Liyan Bai ◽  
...  
Keyword(s):  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Adult Rat ◽  
Adult Rat Hepatocytes

Reappearance and long-term maintenance of connexin32 in proliferated adult rat hepatocytes: use of serum-free L-15 medium supplemented with EGF and DMSO

1995 ◽  
Vol 108 (4) ◽  
pp. 1347-1357
Author(s):  
T. Kojima ◽  
T. Mitaka ◽  
D.L. Paul ◽  
M. Mori ◽  
Y. Mochizuki
Keyword(s):  
Intercellular Communication ◽  
Cell Membranes ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Gap Junctional Intercellular Communication ◽  
Serum Free ◽  
Adult Rat ◽  
Junctional Communication ◽  
Adult Rat Hepatocytes ◽  
Gap Junctional

Intercellular communication, especially gap junctional communication, is thought to be one of the highly differentiated functions of hepatocytes. In primary cultures of rat hepatocytes, it has been considered that the maintenance and the reinduction of differentiated functions is very difficult. In the present study, we succeeded in inducing the gap junctional protein connexin32 (Cx32) in adult rat hepatocytes cultured in serum-free L-15 medium supplemented with epidermal growth factor (EGF) and dimethylsulfoxide (DMSO). When the hepatocytes were cultured in L-15 medium supplemented with 20 mM NaHCO3 and 10 ng/ml EGF in a 5% CO2:95% air incubator, the cells proliferated. Fluorescence immunocytochemistry showed spots immunoreactive to Cx32 on the cell membranes between adjacent cells until day 3, but only a few Cx32-positive spots were found after day 4. Western and northern blot analyses also showed that the amounts of both the protein and mRNA of Cx32 in the cells decreased with time in culture. However, when the cells were treated with 2% DMSO from day 4, the immunoreactive spots reappeared on the cell membranes from day 6 and both their number and intensity gradually increased. The reappearance of Cx32 was accompanied by increases in both the protein and mRNA of Cx32. Furthermore, the expression of Cx32 was well maintained, together with extensive gap junctional intercellular communication, for more than 4 weeks. In addition, ultrastructurally, many gap junctional structures were observed between the hepatocytes, and the antibodies to Cx32 were shown to bind to those structures. This culture system may be useful for studies of the reconstruction of the gap junctional structure, the intracellular pathways of the proteins, and the regulation of synthesis and processing in differentiated hepatocytes.

Receptors for epidermal growth factor (urogastrone) and insulin in primary cultures of rat hepatocytes maintained in serum-free medium

1986 ◽  
Vol 64 (8) ◽  
pp. 803-810 ◽  
Author(s):  
M. O'Connor-McCourt ◽  
M. Soley ◽  
L. J. Hayden ◽  
M. D. Hollenberg
Keyword(s):  
Growth Factor ◽  
Molecular Mass ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Cross Link ◽  
Serum Free ◽  
Epidermal Growth

We have analyzed the receptors for epidermal growth factor (urogastrone) (EGF-URO) and insulin in primary cultures of adult rat hepatocytes maintained for up to 3 weeks on human placental cell matrix in serum-free defined medium. Cross-link labeling experiments revealed that the insulin receptor, partially damaged by the collagenase isolation procedure, was rapidly regenerated to yield an intact receptor. In contrast, cross-link labeling of the EGF-URO receptor revealed that, upon prolonged culture, there was a progressive disappearance of the high molecular mass (175 kilodaltons (kDa)) receptor form, and an appearance of low molecular mass receptor species (130 and 105 kDa). After 3 weeks of culture, the low molecular mass receptor forms accounted for all of the labeled EGF-URO receptor present in the cells. Measurements of EGF-URO binding indicated that the number of EGF-URO binding sites per cell (2.0 × 105 ± 0.3 × 105) did not change during the 3 weeks of culture. However, there was a decrease in EGF-URO binding affinity, reflected by an increase in the KD from 0.6 to 3.0 nM. At zero time and after 3 weeks in culture, Scatchard plots of the binding data were linear; at intermediate time points, the plots were curvilinear. Despite the changes in the EGF-URO receptor that occurred, cells were still responsive to EGF-URO in terms of the inhibition of acetate incorporation into lipid. The ED50 for EGF-URO (about 0.2 nM) was the same for short-term cultures (48 h) as for cells maintained in culture for 3 weeks. We conclude that the long-term culture of hepatocytes in serum-free medium yields an altered low molecular form of the EGF-URO receptor that is, nonetheless, functional. The study points to differential changes in receptors for peptide hormones that may occur in long-term hepatocyte cultures and illustrates the feasibility of using such cultures for metabolic studies of the actions of EGF-URO.

Pyruvate promotion of DNA synthesis in serum-free primary cultures of adult rat hepatocytes

In Vitro ◽  
1983 ◽  
Vol 19 (3) ◽  
pp. 159-166 ◽  
Author(s):  
Joan A. McGowan ◽  
Nancy L. R. Bucher
Keyword(s):  
Dna Synthesis ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Serum Free ◽  
Adult Rat ◽  
Adult Rat Hepatocytes
Sign in / Sign up

Export Citation Format

Share Document