Morphologic and biochemical characteristics of isolated and cultured pancreatic ducts

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

doi:10.1002/1097-0142(19810315)47:6+<1505::aid-cncr2820471411>3.0.co;2-j

Cited by 17 publications

(10 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Since a specific ductal cell marker has yet to be described, morphological and biochemical criteria shown previously to be indicative of ductal cells (7,10,15) were used to characterize the MIA PaCa-2 cell line.…”

supporting

confidence: 93%

“…The predominant cell type comprising the rat (6) and human (7) pancreatic ductal systems are structurally similar throughout the different classes of ducts. Isolation and culture of rat (8)(9)(10) and human pancreatic ducts (Sarras et al, in preparation) have been used previously as a model to study ductal function. Using isolated pancreatic ducts of the pancreatic ductal system have confirmed these findings and have demonstrated that principal cell secretions contain highly sulfated mucins.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Comparative Analysis of a Human Pancreatic Undifferentiated Cell Line (MIA PaCa-2) to Acinar and Ductal Cells

Madden¹,

Heaton²,

Huff³

et al. 1989

Pancreas

View full text Add to dashboard Cite

Pancreatic ductal cell secretion has not been well characterized due to the difficulty in obtaining sufficient quantities of purified ductal cells. To determine if the MIA PaCa-2 cell line would provide a useful model for in vitro studies of pancreatic ductal cell secretion, the present study was designed to characterize these cells in greater detail. In this investigation, the human pancreatic undifferentiated cell line, MIA PaCa-2, was compared with PANC-1 cells (a human ductal cell line previously characterized), isolated rat and human ducts, acinar cells, and nonpancreatic cell lines. The results indicate that while the morphology of the MIA PaCa-2 cell line is nonpolarized and generally atypical of either ductal or acinar cells, the cell line has retained certain biochemical similarities to ductal cells. Additional morphological studies indicated (a) the presence of intermediate filaments characteristic of epithelial cells, (b) the absence of zymogen granules, and (c) an apparent basolateral plasma membrane localization of Na+ , K+-ATPase. Similar to ductal cells, biochemical analyses indicated (a) the presence of N a + , K+-ATPase based on [3H]-ouabain binding assays, (b) high levels of carbonic anhydrase, (c) low levels of y-glutamyl transpeptidase, (d) nondetectable levels of amylase, and (e) protein composition and protein synthetic patterns comparable to PANC-1 cells. Finally, as with PANC-1 cells and isolated rat and human ducts, the major sulfated secretory product of MIA PaCa-2 cells was a protein with a molecular weight of approximately 660,000 to 1 million. These results suggest that while the MIA PaCa-2 cell line can be used to study pancreatic ductal cell secretion, it possesses less ductal-like characteristics than observed with the PANC-1 cell line.

show abstract

supporting

confidence: 93%

mentioning

confidence: 99%

Comparative Analysis of a Human Pancreatic Undifferentiated Cell Line (MIA PaCa-2) to Acinar and Ductal Cells

Madden¹,

Heaton²,

Huff³

et al. 1989

Pancreas

View full text Add to dashboard Cite

show abstract

“…It should be mentioned, however, that as compared with PANC-I cells, significantly higher specific activities were found in both cultured rat and human ducts. This is in agreement with previous studies by Githens et al (29) that indicate that unlike all other enzyme systems monitored, y-glutamyltranspeptidase activity increases with time during culture of rat pancreatic ducts. In contrast, we observed significantly higher specific activities of this enzyme in rat acini, MDCK cells, and L cells.…”

Section: Discussionmentioning

confidence: 89%

“…In comparison with the in situ morphology of the epithelium of rat (M. E. Madden and M. P. Sarras, Jr., in preparation) and human(9) pancreatic ducts, PANC-1 cells grown on a permeable support appear similar to the epithelium observed in the smaller interlobular ducts. As would be expected for an epithelial cell, PANC-1 cells contain cytokeratin, an epithelial class of intermediate filaments.Based on a previous study by Githens et al(29)…”

mentioning

confidence: 90%

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

Madden

Sarras²

1988

Pancreas

View full text Add to dashboard Cite

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.

show abstract

“…This modified digestive procedure both simplified the process and increased the cell yield. In the 1980s, Githens et al (1981Githens et al ( , 1987 and Githens and Whelan (1983) established the explant culture system, which was known as 'classical culture system' and was widely accepted during that period. Nevertheless, explant cultivation takes several weeks, although it is still the most cost-effective method.…”

Section: Discussionmentioning

confidence: 99%

An improved primary culture system of pancreatic duct epithelial cells from Wistar rats

Chen

Zheng

et al. 2009

Cytotechnology

View full text Add to dashboard Cite

Pancreatic duct epithelial cells (PDEC) are involved in most common pancreatic diseases. Primary cultivation of PDEC is a prerequisite for in vitro studies, in which in vivo situations can be simulated and molecular mechanisms investigated better than in cultured cell lines. However, some problems still exist regarding rat PDEC primary cultivation. In this study, an improved primary culture system of rat PDEC is presented. Some modifications, especially regarding specimen chosen, digestive control and epithelium purification, were made to simplify the procedure, increase cell yield, and improve epithelium purification. Cultures were identified as PDEC by morphological characteristics, reverse transcription-polymerase chain reaction and immunocytochemistry staining with cytokeratin 19. In addition, growth characteristics of rat PDEC are described in detail. This improved technique, which is more efficient and cost-effective, will be useful for in vitro pancreatic studies.

show abstract

Morphologic and biochemical characteristics of isolated and cultured pancreatic ducts

Cited by 17 publications

References 18 publications

Comparative Analysis of a Human Pancreatic Undifferentiated Cell Line (MIA PaCa-2) to Acinar and Ductal Cells

Comparative Analysis of a Human Pancreatic Undifferentiated Cell Line (MIA PaCa-2) to Acinar and Ductal Cells

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

An improved primary culture system of pancreatic duct epithelial cells from Wistar rats

Contact Info

Product

Resources

About