Establishment and characterization of chondrocyte cell lines from the costal cartilage of SV40 large T antigen transgenic mice

Murine lung epithelial (MLE) cell lines were produced from lung tumors derived from transgenic mice bearing the viral oncogene, SV40 large T antigen, under transcriptional control of the promoter-enhancer region of the human surfactant protein C (SP-C) gene. Cells were selected on the basis of increased murine cystic fibrosis transmembrane conductance regulator (mCFTR) mRNA content and were dilution cloned to produce distinct immortalized epithelial cell lines. MLE-13a3 cell lines expressing high levels of mCFTR mRNA also expressed apolipoprotein J (apoJ) mRNA, a developmentally regulated glycoprotein expressed preferentially in fetal lung. SP-A, -B, and -C were not detected or were present at low levels in the MLE cells that contained abundant CFTR and apoJ mRNA. In contrast, MLE cells, cloned on the basis of abundant surfactant protein mRNAs, expressed apoJ and mCFTR mRNAs at low levels. Forskolin-stimulated short-circuit current, typical of CFTR-mediated chloride transport activity, was generated by monolayers of subclones of the MLE-13a3 cell lines. Tumor necrosis factor-alpha stimulated mCFTR mRNA, whereas dexamethasone, retinoic acid, and phorbol ester had no effect on the levels of mCFTR mRNA in MLE-13a3 cells.

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Characterization of immortalized human umbilical and iliac vein endothelial cell lines after transfection with SV40 large T-antigen

Blood Coagulation & Fibrinolysis ◽

10.1097/00001721-200001000-00003 ◽

2000 ◽

Vol 11 (1) ◽

pp. 15-25 ◽

Cited By ~ 13

Author(s):

E. B. M. van Leeuwen ◽

R. Veenstra ◽

R. van Wijk ◽

G. Molema ◽

A. Hoekstra ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Lines ◽

Iliac Vein ◽

T Antigen ◽

Large T Antigen ◽

Sv40 Large T Antigen

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Characterization of immortalized human umbilical and iliac vein endothelial cell lines after transfection with SV40 large T-antigen

Blood Coagulation & Fibrinolysis ◽

10.1097/00001721-200011010-00003 ◽

2000 ◽

Vol 11 (1) ◽

pp. 15-25 ◽

Cited By ~ 1

Author(s):

E. B. M. van Leeuwen ◽

R. Veenstra ◽

R. van Wijk ◽

G. Molema ◽

A. Hoekstra ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Lines ◽

Iliac Vein ◽

T Antigen ◽

Large T Antigen ◽

Sv40 Large T Antigen

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Recombinant retroviruses encoding simian virus 40 large T antigen and polyomavirus large and middle T antigens

Molecular and Cellular Biology ◽

10.1128/mcb.6.4.1204-1217.1986 ◽

1986 ◽

Vol 6 (4) ◽

pp. 1204-1217

Author(s):

P S Jat ◽

C L Cepko ◽

R C Mulligan ◽

P A Sharp

Keyword(s):

Cell Lines ◽

Simian Virus 40 ◽

Simian Virus ◽

T Antigen ◽

Vector System ◽

Large T Antigen ◽

T Antigens ◽

Sv40 Large T Antigen ◽

Polyomavirus Middle T Antigen ◽

Middle T Antigen

We used a murine retrovirus shuttle vector system to construct recombinants capable of constitutively expressing the simian virus 40 (SV40) large T antigen and the polyomavirus large and middle T antigens as well as resistance to G418. Subsequently, these recombinants were used to generate cell lines that produced defective helper-free retroviruses carrying each of the viral oncogenes. These recombinant retroviruses were used to analyze the role of the viral genes in transformation of rat F111 cells. Expression of the polyomavirus middle T antigen alone resulted in cell lines that were highly tumorigenic, whereas expression of the polyomavirus large T resulted in cell lines that were highly tumorigenic, whereas expression of the polyomavirus large T resulted in cell lines that were unaltered by the criteria of morphology, anchorage-independent growth, and tumorigenicity. More surprisingly, SV40 large T-expressing cell lines were not tumorigenic despite the fact that they contained elevated levels of cellular p53 and had a high plating efficiency in soft agar. These results suggest that the SV40 large T antigen is not an acute transforming gene like the polyomavirus middle T antigen but is similar to the establishment genes such as myc and adenovirus EIa.

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Growth Properties of Neural Cell Lines Immortalized with the Tsa58 Allele of Sv40 Large T Antigen

Cell Transplantation ◽

10.1177/096368979700600306 ◽

1997 ◽

Vol 6 (3) ◽

pp. 231-238 ◽

Cited By ~ 5

Author(s):

M.E. Truckenmiller ◽

Ora Dillon-Carter ◽

Carlo Tornatore ◽

Henrietta Kulaga ◽

Hidetoshi Takashima ◽

...

Keyword(s):

Amino Acid ◽

Cell Line ◽

Cell Lines ◽

T Antigen ◽

Temperature Sensitive ◽

Permissive Temperature ◽

Large T Antigen ◽

Wild Type ◽

Sv40 Large T Antigen ◽

Growth Properties

In vitro growth properties of three CNS-derived cell lines were compared under a variety of culture conditions. The M213-20 and J30a cell lines were each derived from embryonic CNS culture with the temperature-sensitive (ts) allele of SV40 large T antigen, tsA58, while the A7 cell line was immortalized using wild-type SV40 large T antigen. Cells immortalized with tsA58 SV40 large T proliferate at the permissive temperature, 33° C, while growth is expected to be suppressed at the nonpermissive temperature, 39.5°C. Both the M213-20 and J30a cell lines were capable of proliferating at 39.5°C continuously for up to 6 mo. All three cell lines showed no appreciable differences in growth rates related to temperature over a 7-day period in either serum-containing or defined serum-free media. The percentage of cells in S-phase of the cell cycle did not decrease or was elevated at 39.5°C for all three cell lines. After 3 wk at 39.5°C, the three cell lines also showed positive immunostaining using two monoclonal antibodies reacting with different epitopes of SV40 large T antigen. Double strand DNA sequence analyses of a 300 base pair (bp) fragment of the large T gene from each cell line, which included the ts locus, revealed mutations in both the J30a and M213-20 cell lines. The J30a cell line ts mutation had reverted to wild type, and two additional loci with bp substitutions with predicted amino acid changes were also found. While the ts mutation of the M213-20 cells was retained, an additional bp substitution with a predicted amino acid change was found. The A7 cell line sequence was identical to the reference wild-type sequence. These findings suggest that (a) nucleic acid sequences in the temperature-sensitive region of the tsA58 allele of SV40 large T are not necessarily stable, and (b) temperature sensitivity of cell lines immortalized with tsA58 is not necessarily retained.

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