Immortalization of bovine pancreatic duct epithelial cells

1996 ◽  
Vol 270 (4) ◽  
pp. G676-G683 ◽  
Author(s):  
L. R. Marino ◽  
C. U. Cotton
Keyword(s):  
Epithelial Cells ◽  
Pancreatic Duct ◽  
Cell Line ◽  
Simian Virus 40 ◽  
Duct Cell ◽  
Simian Virus ◽  
T Antigen ◽  
Intracellular Camp ◽  
Large T Antigen ◽  
Pancreatic Duct Cell

Pancreatic duct cell lines have been isolated from a number of animal and human tumors, but none appear to express ion transport properties expected for differentiated pancreatic duct epithelial cells. We sought to generate an immortalized ductal cell line from well-differentiated primary cultures of bovine pancreatic duct epithelium. Epithelial cells from the main duct of the bovine pancreas were isolated and immortalized by transfection with a DNA construct encoding simian virus 40 large T antigen. A single clone (BPD1) survived negative selection and was maintained in culture for > 100 passages over 2 yr. The cells grow readily in culture as monolayers and express several properties characteristic of differentiated pancreatic ductal epithelium. The cells do not appear to form a functional tight junction complex, since the transepithelial resistance of the monolayer cultures grown on a permeable support is < 10 omega.cm2. Northern blot analysis revealed that the cells continue to express simian virus 40 large T antigen and contain significant levels of mRNA for proteins thought to be important in transepithelial bicarbonate secretion [carbonic anhydrase II, Cl-/HCO3- exchanger, Na+/H+ exchanger, and cystic fibrosis transmembrane conductance regulator (CFTR)]. In vivo pancreatic ductal secretion is stimulated by the peptide hormone secretin. The secretin receptor is expressed and functionally coupled to adenylate cyclase in the immortalized cells, since secretin caused a dose-dependent accumulation of adenosine 3'5'-cyclic monophosphate (cAMP; approximately 20-fold increase over basal levels) with a mean effective concentration of 15 nM. Elevation of intracellular cAMP by exposure of the cells to forskolin (10 microM) or secretin (0.1 microM) increase plasma membrane Cl- permeability, most likely mediated by activation of CFTR. The results of these studies demonstrate that the pancreatic duct cell line (BPD1) retains several properties exhibited by the secretory epithelial cells that line the pancreatic ductal tree. This cell line should prove useful for studies of expression, function, and regulation of pancreatic duct cell proteins.

Maintenance of cellular proliferation by adenovirus early region 1A in fibroblasts conditionally immortalized by using simian virus 40 large T antigen requires conserved region 1

1990 ◽  
Vol 10 (12) ◽  
pp. 6664-6673
Author(s):  
T E Riley ◽  
A Follin ◽  
N C Jones ◽  
P S Jat
Keyword(s):  
Cell Line ◽  
Growth Arrest ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Growth Defect ◽  
Primary Cells ◽  
Nonpermissive Temperature ◽  
Large T Antigen ◽  
Adenovirus E1a

Various mutants of adenovirus E1A were assayed for their ability to complement the growth defect at the nonpermissive temperature for the cell line tsa14 which was isolated by immortalizing rat embryo fibroblasts with the thermolabile large T antigen of tsA58. This cell line grows indefinitely at the permissive temperature but undergoes rapid growth arrest upon shift up to the nonpermissive temperature. Since this growth arrest can be overcome by introduction of wild-type simian virus 40 large T antigen, human papillomavirus 16 E7, and adenovirus E1A, the tsa14 cells provided an excellent system for defining regions of E1A necessary for complementation of the growth defect. We demonstrate that conserved region 1 (CR1) is the region of E1A required for complementation. While CR2 of E1A has been shown to be required for the immortalization of primary cells and is also necessary for the binding of the 105-kDa retinoblastoma protein, mutations within this region did not abrogate complementation of the growth defect. However, since both CR1 and CR2 have previously been shown to be absolutely required for immortalization of primary cells by adenovirus E1A, this evidence suggests that the tsa14 system assays for the maintenance of proliferation and that this requires CR1.

Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells

1987 ◽  
Vol 7 (5) ◽  
pp. 2031-2034
Author(s):  
D E Brash ◽  
R R Reddel ◽  
M Quanrud ◽  
K Yang ◽  
M P Farrell ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Primary Culture ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Bronchial Epithelial Cells ◽  
Large T Antigen ◽  
Bronchial Epithelial ◽  
Antigen Gene ◽  
Strontium Phosphate

Strontium ion formed DNA-phosphate precipitates analogous to those formed by calcium but lacking the lethal and differentiation-inducing effects of calcium on many epithelial cell types in primary culture. Human primary bronchial epithelial cells were transiently and stably transfected by using strontium phosphate; the frequency of stable transformation with a plasmid carrying the simian virus 40 large-T-antigen gene was greater than 10(-4).

scholarly journals Maintenance of cellular proliferation by adenovirus early region 1A in fibroblasts conditionally immortalized by using simian virus 40 large T antigen requires conserved region 1.

1990 ◽  
Vol 10 (12) ◽  
pp. 6664-6673 ◽  
Author(s):  
T E Riley ◽  
A Follin ◽  
N C Jones ◽  
P S Jat
Keyword(s):  
Cell Line ◽  
Growth Arrest ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Growth Defect ◽  
Primary Cells ◽  
Nonpermissive Temperature ◽  
Large T Antigen ◽  
Adenovirus E1a

Various mutants of adenovirus E1A were assayed for their ability to complement the growth defect at the nonpermissive temperature for the cell line tsa14 which was isolated by immortalizing rat embryo fibroblasts with the thermolabile large T antigen of tsA58. This cell line grows indefinitely at the permissive temperature but undergoes rapid growth arrest upon shift up to the nonpermissive temperature. Since this growth arrest can be overcome by introduction of wild-type simian virus 40 large T antigen, human papillomavirus 16 E7, and adenovirus E1A, the tsa14 cells provided an excellent system for defining regions of E1A necessary for complementation of the growth defect. We demonstrate that conserved region 1 (CR1) is the region of E1A required for complementation. While CR2 of E1A has been shown to be required for the immortalization of primary cells and is also necessary for the binding of the 105-kDa retinoblastoma protein, mutations within this region did not abrogate complementation of the growth defect. However, since both CR1 and CR2 have previously been shown to be absolutely required for immortalization of primary cells by adenovirus E1A, this evidence suggests that the tsa14 system assays for the maintenance of proliferation and that this requires CR1.

Immortalization of Rat Eustachian Tube Epithelial Cells by Adenovirus 12-Simian Virus 40 Hybrid Virus

2002 ◽  
Vol 111 (10) ◽  
pp. 919-925 ◽  
Author(s):  
Sunji Jin ◽  
Shigehiro Ueyama ◽  
Sung-Kyun Moon ◽  
Johng S. Rhim ◽  
Xin-Xing Gu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Line ◽  
Otitis Media ◽  
Eustachian Tube ◽  
Primary Cultures ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Male Rat ◽  
Positive Staining

The eustachian tube epithelial cells play an important role in the initial pathogenesis of otitis media. In order to study the role of the eustachian tube epithelial cells in the pathogenesis of otitis media, we have established a rat eustachian tube epithelial cell line. The cell line was derived by infecting primary cultures of eustachian tube epithelial cells with the adenovirus 12-simian virus 40 (Ad12-SV40) hybrid virus. The immortalized cells have retained the morphological characteristics of the parental cells and show positive staining with anti-cytokeratin antibodies (a marker for epithelial cells), but not with anti-vimentin antibodies (a fibroblast marker). The cells have been in continuous culture for more than 10 months and have undergone 38 passages. Western blotting and cell staining have confirmed the expression of the SV40 T antigen and p53. Chromosomal analysis indicates that the cell line is aneuploid and derived from male rat epithelial cells. Together, our results suggest that the cell line originated from eustachian tube epithelial cells from a male rat and was successfully immortalized by the Ad12-SV40 virus.

Establishment of a novel chondrocyte-like cell line derived from transgenic mice harboring the temperature-sensitive simian virus 40 large T-antigen gene

2009 ◽  
Vol 11 (11) ◽  
pp. 1646-1654 ◽  
Author(s):  
Nobuko Mataga ◽  
Masato Tamura ◽  
Nobuyuki Yanai ◽  
Tamayuki Shinomura ◽  
Koji Kimata ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Cell Line ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Temperature Sensitive ◽  
Large T Antigen ◽  
Antigen Gene

scholarly journals Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells.

1987 ◽  
Vol 7 (5) ◽  
pp. 2031-2034 ◽  
Author(s):  
D E Brash ◽  
R R Reddel ◽  
M Quanrud ◽  
K Yang ◽  
M P Farrell ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Primary Culture ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Bronchial Epithelial Cells ◽  
Large T Antigen ◽  
Bronchial Epithelial ◽  
Antigen Gene ◽  
Strontium Phosphate

Strontium ion formed DNA-phosphate precipitates analogous to those formed by calcium but lacking the lethal and differentiation-inducing effects of calcium on many epithelial cell types in primary culture. Human primary bronchial epithelial cells were transiently and stably transfected by using strontium phosphate; the frequency of stable transformation with a plasmid carrying the simian virus 40 large-T-antigen gene was greater than 10(-4).

scholarly journals Analysis of a large-T-antigen variant expressed in simian virus 40-transformed mouse cell line mKS-A.

1989 ◽  
Vol 63 (6) ◽  
pp. 2820-2828 ◽  
Author(s):  
S Dora ◽  
C Schwarz ◽  
M Baack ◽  
A Graessmann ◽  
R Knippers
Keyword(s):  
Cell Line ◽  
Simian Virus 40 ◽  
Mouse Cell ◽  
Simian Virus ◽  
T Antigen ◽  
Large T Antigen ◽  
Mouse Cell Line ◽  
Antigen Variant
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