Immortalization of human corneal epithelial cells using simian virus 40 large T antigen and cell characterization

2016 ◽  
Vol 78 ◽  
pp. 52-57 ◽  
Author(s):  
Cho-Won Kim ◽  
Ryeo-Eun Go ◽  
Geum-A Lee ◽  
Chang Deok Kim ◽  
Young-Jin Chun ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Large T Antigen ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Cell Characterization ◽  
Human Corneal Epithelial Cells

scholarly journals The transcriptome of wild-type and immortalized corneal epithelial cells

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Kai Furuya ◽  
Tao Wu ◽  
Ai Orimoto ◽  
Eriko Sugano ◽  
Hiroshi Tomita ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Expression Pattern ◽  
Cultured Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Cyclin Dependent Kinase ◽  
Wild Type ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial

AbstractCellular immortalization enables indefinite expansion of cultured cells. However, the process of cell immortalization sometimes changes the original nature of primary cells. In this study, we performed expression profiling of poly A-tailed RNA from primary and immortalized corneal epithelial cells expressing Simian virus 40 large T antigen (SV40) or the combination of mutant cyclin-dependent kinase 4 (CDK4), cyclin D1, and telomere reverse transcriptase (TERT). Furthermore, we studied the expression profile of SV40 cells cultured in medium with or without serum. The profiling of whole expression pattern revealed that immortalized corneal epithelial cells with SV40 showed a distinct expression pattern from wild-type cells regardless of the presence or absence of serum, while corneal epithelial cells with combinatorial expression showed an expression pattern relatively closer to that of wild-type cells.

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 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 Unexpected thymic hyperplasia in transgenic mice harboring a neuronal promoter fused with simian virus 40 large T antigen.

1987 ◽  
Vol 7 (9) ◽  
pp. 3178-3184 ◽  
Author(s):  
F M Botteri ◽  
H van der Putten ◽  
D F Wong ◽  
C A Sauvage ◽  
R M Evans
Keyword(s):  
Growth Hormone ◽  
Epithelial Cells ◽  
Transgenic Mice ◽  
Simian Virus 40 ◽  
Immunohistochemical Analysis ◽  
Simian Virus ◽  
T Antigen ◽  
Thymic Hyperplasia ◽  
Thymic Epithelial Cells ◽  
Large T Antigen

The hypothalamic peptide growth hormone-releasing factor (GRF) regulates the secretion and production of growth hormone from the anterior pituitary (M. C. Gelato and G. R. Merriam, Annu. Rev. Physiol. 48:569-591). To study GRF gene regulation, transgenic mice were generated that harbor the human GRF promoter fused to the coding sequences from the simian virus 40 early region. These mice had normal hypothalamic functions but unexpectedly suffered from severe thymic hyperplasia. Immunohistochemical analysis revealed that large T antigen was expressed in the thymic epithelial cells. These cells have endocrine properties and are known to produce thymic hormones [corrected]. The thymic hyperplasia was the apparent consequence of inappropriate production of T-cell maturation factors by epithelial cells and could involve increased self renewal of apparently normal T stem cells in the thymus.

Unexpected thymic hyperplasia in transgenic mice harboring a neuronal promoter fused with simian virus 40 large T antigen

1987 ◽  
Vol 7 (9) ◽  
pp. 3178-3184
Author(s):  
F M Botteri ◽  
H van der Putten ◽  
D F Wong ◽  
C A Sauvage ◽  
R M Evans
Keyword(s):  
Growth Hormone ◽  
Epithelial Cells ◽  
Transgenic Mice ◽  
Simian Virus 40 ◽  
Immunohistochemical Analysis ◽  
Simian Virus ◽  
T Antigen ◽  
Thymic Hyperplasia ◽  
Thymic Epithelial Cells ◽  
Large T Antigen

The hypothalamic peptide growth hormone-releasing factor (GRF) regulates the secretion and production of growth hormone from the anterior pituitary (M. C. Gelato and G. R. Merriam, Annu. Rev. Physiol. 48:569-591). To study GRF gene regulation, transgenic mice were generated that harbor the human GRF promoter fused to the coding sequences from the simian virus 40 early region. These mice had normal hypothalamic functions but unexpectedly suffered from severe thymic hyperplasia. Immunohistochemical analysis revealed that large T antigen was expressed in the thymic epithelial cells. These cells have endocrine properties and are known to produce thymic hormones [corrected]. The thymic hyperplasia was the apparent consequence of inappropriate production of T-cell maturation factors by epithelial cells and could involve increased self renewal of apparently normal T stem cells in the thymus.

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.

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