A Method of Maintaining Parenchymal Cells from Adult Rat Liver In Vitro

1972 ◽  
Vol 11 (1) ◽  
pp. 249-260
Author(s):  
J. ALWEN ◽  
JENNIFER J. GALLHAI-ATCHARD
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Protein Synthesis ◽  
Light And Electron Microscopy ◽  
Rat Hepatocytes ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Rna And Protein Synthesis ◽  
Parenchymal Cells

A method for preparing suspensions of adult rat hepatocytes suitable for maintenance in vitro is described. Cultures were established from the cell suspensions by the squash technique. Cells were examined by light and electron microscopy; histochemically for glycogen, bile, lipid and glucose-6-phosphatase; and by autoradiography for DNA, RNA and protein synthesis. Hepatocytes could be maintained in vitro for at least 3 days and began to aggregate after 1 day. Uridine and leucine were incorporated, but not thymidine. Cultures consisted mainly of hepatocytes, though reticulo-endothelial cells were sometimes present.

Immunoenzymatic demonstration of the simultaneous presence of transferrin, hemopexin and albumin in a same hepatocyte and their ultrastructural localization

1978 ◽  
Vol 36 (2) ◽  
pp. 88-89
Author(s):  
M. Kraemer ◽  
J. Foucrier ◽  
J. Vassy ◽  
M.T. Chalumeau
Keyword(s):  
Plasma Proteins ◽  
Rat Hepatocytes ◽  
Ultrastructural Localization ◽  
Carrier Proteins ◽  
Normal Cells ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Monospecific Antisera ◽  
Different Levels

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.

scholarly journals The effect of insulin-dextran complexes on the protein synthesis in the primary monolayer culture of adult rat hepatocytes.

1984 ◽  
Vol 144 (3) ◽  
pp. 245-256
Author(s):  
TSUGUHIKO NAKAI ◽  
YASUNORI KUTSUMI ◽  
YOSHIKAZU SAKAMOTO ◽  
KOJI OIDA ◽  
SUSUMU MIYABO ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Monolayer Culture ◽  
Rat Hepatocytes ◽  
Adult Rat ◽  
Primary Monolayer Culture ◽  
Adult Rat Hepatocytes ◽  
Primary Monolayer

Cellular actin and junction formation during reaggregation of adult rat hepatocytes into epithelial cell sheets

1978 ◽  
Vol 31 (1) ◽  
pp. 341-353
Author(s):  
A. Miettinen ◽  
I. Virtanen ◽  
E. Linder
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cell ◽  
Indirect Immunofluorescence ◽  
Rat Hepatocytes ◽  
Immunofluorescence Technique ◽  
Cell Sheets ◽  
Adult Rat ◽  
Junction Formation ◽  
Adult Rat Hepatocytes ◽  
Indirect Immunofluorescence Technique

Aggregation of adult rat hepatocytes, isolated by the collagenase perfusion technique, was studied by ultrastructural methods and the indirect immunofluorescence technique using anti-actin antibodies. In a primary culture the cells rapidly made contact with each other by filopodia-like structures, as seen by scanning electron microscopy. In a few hours this led to stable adhesion of the cells. No identifiable junction formation occurred during the first 17 h in culture. Within 48 h the cells had formed epithelial cell sheets with junctional complexes consisting of tight junctions, bile canaliculus-like structures and zonula adhaerens-type junctions. The distribution of cytoplasmic actin fluorescence remained homogenous in the contacting cells during the first 24 h in culture, as seen with anti-actin antibodies by the indirect immunofluorescence technique. The first short, fluorescent actin filaments appeared in the periphery of the developing lamellipodia of the spreading cell islands. In organized epithelial cell sheets these filaments were seen as long fibres ending at the perinuclear region of the marginal cells. In the submarginal cells fluorescent actin fibres were distinct at the junctional regions of the cells. This filamentous fluorescence seemed to extend throughout the entire cell sheet in an organized manner and correspond to the apical layer of parallel microfilaments seen in transmission electron microscopy. Our results suggest that filamentous actin plays a role in the contact-induced spreading of the cells and in the maintenance of the internal organization of the epithelial cell sheets.

scholarly journals Ultrastructural indirect immunolocalization of transferrin in cultured rat hepatocytes permeabilized with saponin.

1984 ◽  
Vol 32 (5) ◽  
pp. 538-540 ◽  
Author(s):  
J Vassy ◽  
M Rissel ◽  
M Kraemer ◽  
J Foucrier ◽  
A Guillouzo
Keyword(s):  
Endoplasmic Reticulum ◽  
Intact Cell ◽  
Rat Hepatocytes ◽  
Cell Types ◽  
Glutaraldehyde Fixation ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Cultured Rat Hepatocytes ◽  
Indirect Immunoperoxidase ◽  
Parenchymal Cells

Transferrin was localized in 48-hr cultured adult rat hepatocytes by indirect immunoperoxidase following paraformaldehyde--glutaraldehyde fixation and the use of saponin as a membrane permeabilizing agent. The protein, present in all the parenchymal cells in variable amounts, was found to be specifically located in the endoplasmic reticulum and Golgi apparatus. These results are consistent with recent reports claiming that all adult hepatocytes may synthesize a given liver plasma protein at a given time. The procedure used in this study should be particularly useful for the detection of intracellular antigens in various intact cell types.

scholarly journals Hormonal regulation of glutathione S-transferase expression in co-cultured adult rat hepatocytes

2000 ◽  
Vol 166 (2) ◽  
pp. 363-371 ◽  
Author(s):  
S Coecke ◽  
T Vanhaecke ◽  
A Foriers ◽  
IR Phillips ◽  
A Vercruysse ◽  
...  
Keyword(s):  
Hormonal Regulation ◽  
Cultured Cells ◽  
Experimental System ◽  
Human Growth ◽  
Rat Hepatocytes ◽  
Isolated Hepatocytes ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Translational Level

Glutathione S-transferases (GSTs) are subject to regulation by thyroid and sex hormones and by GH. We have used an in vitro experimental system comprising adult rat hepatocytes co-cultured with rat liver epithelial cells of primitive biliary origin, to distinguish between direct and indirect effects of various hormones on GSTs; to identify the GST subunits affected by individual hormones; and to investigate the level at which the hormones act. Tri-iodothyronine (T3), thyroxine (T4) and 17beta-oestradiol (OE2) reduced GST activities, whereas testosterone, dihydrotestosterone, and human growth hormone (hGH) had little effect on total GST activity. HPLC separation of the various GST subunits revealed that T3 and T4 reduced total GST content, in particular the abundance of subunits M1 and M2. The amount of the Pi-class subunit P1 was reduced by OE2. Treatment of the co-cultured cells with this hormone altered the GST subunit profile to one that is more similar to that observed in freshly isolated hepatocytes. Analysis of mRNAs demonstrated that some of the hormones act at a pre-translational level, whereas others act at a translational or post-translational level to regulate the expression of various GST subunits.

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