Cultures from Adult Rat Liver Cells. II. Demonstration of Organ-Specific Cell Surface Antigens on Cultured Cells from Normal Liver

Iype, P T; Baldwin, Robert; Glaves, Dorothy

doi:10.1038/bjc.1972.3

Cited by 28 publications

(9 citation statements)

References 8 publications

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“…These cell-lines, which had been in cell culture for 120-150 days were stored frozen before use in these experiments. These control cells have been shown to possess certain antigenic (Iype et al, 1972), morphological Allen et a1., 1978), and biochemical , characteristics of rat liver and did not possess any markers associated with hepatoma cells (Iype and Allen, 1979). The liver cell strain LNRL 1 was isolated from a 13-day-old male Lewis rat by enzymatic digestion using a mixture of Collagenase/Dispase (Boehringer Mannheim Biochemicals, Indianapolis, Indiana).…”

Section: Cell Lines and Culture Methodsmentioning

confidence: 99%

In Vitro Toxicity Testing of Environmental Agents

Kolber¹,

Wong²,

Grant³

et al. 1983

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Section: Cell Lines and Culture Methodsmentioning

confidence: 99%

In Vitro Toxicity Testing of Environmental Agents

Kolber¹,

Wong²,

Grant³

et al. 1983

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“…Since that time they have been characterized with respect to karyotype (Chessebeuf et al, 1981;Tsao et al, 1985a), ultrastructure Grisham, 1975;Furukawa et al, 1987), intermediate filaments (Mayer and SchMer, 1982;Marceau et al, 1986;Neupert et al, 1987), membrane antigens (Iype et al, 1972;Marceau et al, 1986), enzyme content Mayer and Schkfer, 1982), amino acid transport (Gebhardt and Williams, 1986) and various cellular and metabolic functions (Tsao et al, 1984;Ismail-Beigi et al, 1986). Despite the considerable variations in phenotypic properties of the different cell lines revealed by these studies, the lines have proved useful for investigating many aspects of liver function, including enzyme regulation (Gerschenson et al, 1974) and metabolism of xenobiotics (Williams, 1981).…”

Section: Introductionmentioning

confidence: 99%

“…Although the epithelial cell lines have maintained some of the functional characteristics of hepatocytes (Borek, 1972;Iype et al, 1972;Weinstein et al, 1975;Odashima et al, 1976;Mayer and Sch~fer, 1982;Herring et al, 1983;Gebhardt and Williams, 1986), there is substantial evidence that the epithelial cells are not derived from hepatocytes (Grisham et al, 1975;Furukawa et al, 1987). However, the exact origin of these cells in the liver is still unresolved.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibodies directed against rat liver epithelial cell lines selectively recognize bile duct epithelium in livers of adult rats

Gebhardt

Schäfer-Degenhart

1988

Cell Biol Toxicol

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Monoclonal antibodies directed against antigens on rat liver epithelial cell lines were prepared. Three antibodies, 4C3, 19C6, and 3C2, recognized surface antigens present (although in different quantities) on eight epithelial cell lines tested, irrespective of whether they were normal or transformed. For MAb 3C2, the primary antigen common to all but one cell line showed a Mr of 135 kD. In paraffin sections of liver tissue, two antibodies, 4C3 and 19C6, reacted exclusively with bile duct epithelium, whereas the MAb 3C2 additionally reacted with sinusoidal endothelium and the endothelium of the portal venules. In sections of livers from rats exposed to diethylnitrosamine, the MAb 19C6 selectively stained bile duct-like structures in cholangiomas, while other preneoplastic and neoplastic lesions were not stained. These results demonstrate that the monoclonal antibodies obtained may prove useful for investigating cell lineages related to propagable liver epithelial cell lines and suggest that these cells may be derived from terminal bile ductular cells.

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Characterization of Gluconeogenesis in Enzymatically Isolated Parenchymal Cells of Rat Liver

Cornell

Filkins

1974

Experimental Biology and Medicine

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Evaluation of hepatic paren'chyma.1 cell metabolic functions in vitro has been attempted utilizing a number of test systems, viz., the perfused liver (1, 2) liver slices (3, 4), andl liver cells isolated by a variety of techniques (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15). While studies of the perfused rat liver have been extensive and successful, two1 major disadvantages of this system are (i) it is not exclusively a population of parenchymal cells, and (ii) the number of variables able to be studied simultaneously is limited. The value of liver slices is also restricted since they are a nonuniform and complex system in which oxygen diffusion to cells on the inside of the slice is limited and cells on the outer surface are mechanically damaged. A homogeneous preparation od intact parenchymal cells overcomes many of the disadvantages mentioned in the other systems and, in addition, permits the study of many variables while also providing simultaneous control samples.The purpose of the present investigation was to characterize a method for the preparation of intact viable hepatocytes with particular emphasis on their application to the in ;Jitro study of gluconeogenesis. The method employed was a modification of the enzymatic technique developed by Berry and Friend (6) and involved perfusion of isolated rat livers with media containing collagenase and hyaluronidase. The ability of the parenchymal cells isolated by this method to exclude vital dyes and to retain enzymes localized to the cytoplasmic, mitochondrial, and lysosomal portions of the cell was deter-mined. These tests were deemed critical because mechanically isolated hepatocytes have been shown to be demaged sufficiently to take up trypan blue and to leak cytoplasmfic enzymles (5, 9, 11). Since Krebs and coworkers ( 2 ) have stated that a stringent test of the metabolic integrity of the hepatocyte is its ability to synthesize glucose from lactate, the gluconeogenic capacity of the isolated cells was evaluated in the present study. Of special interest was the inlfluence of fasting duration on hepatocyte gluconeogenesis in light of recent evidence identifying peripheral substrate release as the probable rate controllling factor in hepatic gluconeogenesis during prolonged !fasting (116). Materials and Methods. A n h a b .Male rats of the Holtzman strain (Holtzman Co., Madison, WI) weighing 280-310 g were fed Purina chow and water ad libitum in animal quarters maintained a t 76-78' F and at an automatically regulated 12-hr light-dark cycle (7 AM-7 P M ) . Experiments commenced a t 9 AM on rats which were fasted with access to water for 24, 48, 96, or 168 hr prior to use.Liver isolation. Parenchymal cells were isolated from livers by a modification of the method of Berry and Frliend (6). Rats were anesthetized with sodium pentobarbital a t a dose of 30 mg/kg via the dorsal vein of the penis. A midline laporotomy was performed, the inferior vena cava and portal vein were isolated, and 500 USP units of heparin were injected iv. After ligating the abdominal vena ...

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Cultures from Adult Rat Liver Cells. II. Demonstration of Organ-Specific Cell Surface Antigens on Cultured Cells from Normal Liver

Cited by 28 publications

References 8 publications

In Vitro Toxicity Testing of Environmental Agents

In Vitro Toxicity Testing of Environmental Agents

Monoclonal antibodies directed against rat liver epithelial cell lines selectively recognize bile duct epithelium in livers of adult rats

Characterization of Gluconeogenesis in Enzymatically Isolated Parenchymal Cells of Rat Liver

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