Ducts of the rat pancreas in agarose matrix culture

Githens, Sherwood; Holmquist, Doris R. G.; Whelan, Joelle F.; Ruby, John R.

doi:10.1007/bf02619315

Cited by 27 publications

(13 citation statements)

References 19 publications

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“…Information gained from other studies provides an additional reasonable explanation for lumen enlargement. The ends of pancreatic ducts in culture seal off to create enclosed lumens (Githens et al, 1980). As a result the lumens enlarge and the epithelial cells flatten at the periphery.…”

Section: Enlargement Of Lumensmentioning

confidence: 99%

Origin and Development of the Precursor Lesions in Experimental Pancreatic Cancer in Rats

Bockman

Guo

Büchler

et al. 2003

Laboratory Investigation

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SUMMARY:Notwithstanding the importance of understanding how pancreatic ductal adenocarcinoma develops, the process remains controversial. A key question is whether the cells of origin of the tubular complexes that constitute precursor lesions are derived from a single cell type or from multiple types. Suggestions that they arise solely from centroacinar cells or ductal cells have been based on inference due to their morphologic appearance in tissue from patients or investigation of limited numbers of tubular complexes in animal models later in the carcinogenic process. The present study establishes clearly that two steps are involved; rapid transdifferentiation to produce tubular complexes followed later by transformation of the component cells. Animals were killed at intervals beginning 1 day after implantation of the carcinogen dimethylbenzanthracene. Transdifferentiation of acinar cells to ductal cells does not require cell division. Transition of lobules to tubular complexes begins by 2 days after implantation of carcinogen. Within 4 days after implantation well-developed tubular complexes are present. Islets participate in the process. Ductal adenocarcinoma is observed by 1 month after implantation of carcinogen. Chymotrypsin and cytokeratin localized by immunocytochemistry indicate acinar and ductal cell characteristics. Acino-ductal transdifferentiation persists in carcinogen-implanted animals, but not in controls implanted with sodium chloride crystals or subjected to sham implantation. The precursor lesions (tubular complexes) are formed by the transdifferentiation of acinar cells and to a lesser extent islet cells, with the incorporation of the duct cells pre-existing in the lobules. Therefore, cells that at one time were acinar cells, islet cells, and duct cells, provide the precursor cells for the ductal adenocarcinoma that develops from tubular complexes. The results raise the question whether the transdifferentiated cells in the tubular complexes of patients with chronic pancreatitis are more susceptible to carcinogenic influences, resulting in the increased rate of pancreatic cancer. (Lab Invest 2003, 83:853-859).

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Section: Enlargement Of Lumensmentioning

confidence: 99%

Origin and Development of the Precursor Lesions in Experimental Pancreatic Cancer in Rats

Bockman

Guo

Büchler

et al. 2003

Laboratory Investigation

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“…There were twelve ducts in each group and the vertical and horizontal bars represent the S.E.M. DISCUSSION Githens, Holmquist, Whelan & Ruby (1980) first showed that ducts isolated from the pancreas of copper-replete rats sealed within 2-4 days in culture. We attribute the more rapid sealing (within 8 h) of ducts isolated from copper-deficient animals to their excellent morphological and biochemical preservation (Arkle et al 1986).…”

Section: Permeability Of Ductal Epitheliummentioning

confidence: 99%

Morphological, Biochemical and Secretory Studies on Rat Pancreatic Ducts Maintained in Tissue Culture

Argent¹,

et al. 1986

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SUMMARYInterlobular ducts were isolated from the pancreas of copper-deficient rats and maintained in culture on polycarbonate filter rafts. Within 8 h the ends of the ducts had sealed. This was followed by a marked dilatation of the lumen, a flattening of the epithelium against the surrounding connective tissue layer and an over-all swelling of the duct. Apart from a reduction in their height, a fall in the number of intracellular fat droplets and a widening of intercellular spaces, epithelial cells within the cultured ducts retained all the ultrastructural characteristics of those in freshly isolated preparations. The basal concentration of adenosine 3',5'-phosphate (cyclic AMP) in the cultured ducts was 43 3 +6 8 ,umol .1-1 duct epithelium (n = 5) and was increased to 1889 + 55-2 ,umol . 1-1 duct epithelium (n = 5) in the presence of 10-8 M secretin. The basal rate of fluid secretion, measured using micropuncture techniques, was 0-16 + 0 03 nl. h-. nl-l duct epithelium (n = 12). This was increased 14-fold by 10-8 M secretin while the dose of the hormone required for half-maximal secretion was about 2 x 10-11 mol . 1-1. The concentration of chloride ions in secreted fluid and perifusion buffer were similar. Variation in culture time up to 52 h had no effect on fluid secretion, and the response to secretin was dependent on the presence of bicarbonate ions in the perifusion fluid. N6,02-dibutyryl adenosine 3',5'-phosphate (dibutyryl cyclic AMP) also increased fluid secretion but caerulein had no effect. We suggest that isolated ducts secrete fluid at comparable rates to ducts in situ within the pancreas of copper-replete rats.

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“…The isolated fragments were cultured in RPMI 1640-SeaPrep medium as described by Githens et al (10). The ducts seal within I day of culture and begin to balloon while the acinar cells become necrotic.…”

Section: Cell Line and Culture Conditionsmentioning

confidence: 99%

“…To further analyze if the discharge of PANC-1 sulfated products was a bona fide secretory process and not just cell leakage, the cells were pulsed for 10 min in KRH with 35S04 and then chased for 2 h in KRH + SO, at 4°C or in the presence of energy inhibitors such as rotenone (10 pg/ml) or oligomycin (20 pg/ml) dissolved in 2% ETOH. As shown in Fig.…”

Section: Biochemical Characterizationmentioning

confidence: 99%

Morphological and Biochemical Characterization of a Human Pancreatic Ductal Cell Line (PANC- 1)

Madden

Sarras²

1988

Pancreas

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Study of the products secreted by pancreatic ductal cells and analysis of the mechanisms involved in the discharge of these products have been limited by a lack of in vitro models available to experimentally approach this problem. To this aim, this investigation has been designed to determine if a human pancreatic carcinoma cell line of ductal origin (PANC-1) has maintained some of the differentiated characteristics of normal mammalian pancreatic ductal epithelium. Morphological and immunocytochemical studies indicated that, similar to isolated rat pancreatic ducts, the PANC-1 cell line contained (a) intermediate filaments of the epithelial class, (b) a basolateral plasma membrane localization of N a + , KC-ATPase, (c) complete tight junctions based on freeze-fracture analysis, (d) a cuboidal morphology when grown on Type I collagen-coated nitrocellulose filters or isolated amnion basement membrane, and (e) normal ductal epithelial ultrastructural features. Biochemical analysis indicated that, also similar to isolated rat and human pancreatic ducts, the PANC-1 cell line contained (a) y-glutamyltranspeptidase, (b) carbonic anhydrase, and (c) N a + , K+-ATPase based on [3H]ouabain binding assays. Comparative studies with other transformed lines indicated that PANC-1 cells have similarities to ductal lines such as MDCK cells but are markedly different from mesenchymally derived lines such as L cells. In addition, as with isolated rat and human ducts, PANC-1 cells synthesize and secrete sulfated proteins with a MW range of -180K to 1 million daltons, with the predominant species being 660K daltons as indicated by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis. These results indicate that the PANC-1 cell line has maintained at least some of the differentiated characteristics of normal pancreatic ductal epithelial cells and may be a useful system for study of ductal secretory products as well as the mechanisms involved in the discharge of these products.

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Ducts of the rat pancreas in agarose matrix culture

Cited by 27 publications

References 19 publications

Origin and Development of the Precursor Lesions in Experimental Pancreatic Cancer in Rats

Origin and Development of the Precursor Lesions in Experimental Pancreatic Cancer in Rats

Morphological, Biochemical and Secretory Studies on Rat Pancreatic Ducts Maintained in Tissue Culture

Morphological and Biochemical Characterization of a Human Pancreatic Ductal Cell Line (PANC- 1)

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