Immortalization of male genital duct epithelium: an assay system for the cystic fibrosis gene

1991 ◽  
Vol 98 (1) ◽  
pp. 85-89
Author(s):  
L. Coleman ◽  
A. Harris
Keyword(s):  
Cystic Fibrosis ◽  
Epithelial Cells ◽  
Autosomal Recessive ◽  
Vas Deferens ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Cystic Fibrosis Gene ◽  
Duct Epithelium ◽  
Male Genital

The epithelia lining the vas deferens and epididymis are directly involved in the pathology of the autosomal recessive disease cystic fibrosis (CF). We have established culture systems for these epithelial cells. Long-term cell lines have now been generated from these primary epithelial cells by transformation with a plasmid containing an origin-defective simian virus 40 (SV40). Lines have been established from vas deferens and epididymis and both maintain expression of the CF gene.

CONGENITAL ABSENCE OF THE VAS DEFERENS: INCOMPLETE PENETRANCE OF CYSTIC FIBROSIS GENE MUTATIONS

1997 ◽  
Vol 158 (5) ◽  
pp. 1794-1799 ◽  
Author(s):  
David Shin ◽  
Fred Gilbert ◽  
Marc Goldstein ◽  
Peter N. Schlegel
Keyword(s):  
Cystic Fibrosis ◽  
Vas Deferens ◽  
Gene Mutations ◽  
Congenital Absence ◽  
Cystic Fibrosis Gene ◽  
Incomplete Penetrance

Cystic Fibrosis Manifested as Undescended Testis and Absence of Vas Deferens

PEDIATRICS ◽  
1992 ◽  
Vol 90 (6) ◽  
pp. 982-983
Author(s):  
RAN GOSHEN ◽  
EITAN KEREM ◽  
TZIPORA SHOSHANI ◽  
BAT-SHEVA KEREM ◽  
ELAD FEIGIN ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Autosomal Recessive ◽  
Undescended Testis ◽  
Vas Deferens ◽  
Pancreatic Insufficiency ◽  
European Ancestry ◽  
Inherited Disease ◽  
Musculoskeletal Problems ◽  
Male Patients ◽  
Almost All

Cystic fibrosis (CF) is the most common autosomal recessive inherited disease in whites. Among whites of European ancestry, approximately 1 in 2000 live births are affected, implying a carrier frequency of 1:25.1 The disease is characterized by chronic lung disease, which usually leads to the patient's death. Furthermore, patients with CF suffer from pancreatic insufficiency and other less common manifestations, such as meconium ileus, hepatobiliary abnormalities, diabetes mellitus, and musculoskeletal problems.1 Almost all males with CF are infertile. Absence of the vas deferens has been reported in 70% to more than 90% of male patients affected by CF.2 Although 4% of full-term male neonates have un-descended testes at birth, 0.8% to 1.0% of 1-year-old boys have cryptorchism and may be subjected to a later surgical intervention.3

Effect of 5-azacytidine on gene expression in marrow stromal cells

1989 ◽  
Vol 9 (6) ◽  
pp. 2748-2751
Author(s):  
D F Andrews ◽  
J Nemunaitis ◽  
C Tompkins ◽  
J W Singer
Keyword(s):  
Stromal Cells ◽  
Time Course ◽  
Matrix Protein ◽  
Interleukin 1 ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Marrow Stromal Cells ◽  
Necrosis Factor Alpha ◽  
Factor Alpha

When exposed to 5-azacytidine, marrow stromal cells from active long-term marrow cultures and cell lines derived from simian virus 40-transformed stromal cells rapidly upregulated c-abl and interleukin-6 transcripts while downregulating the expression of collagen I, a major matrix protein. Similar effects occurred with interleukin-1 alpha and tumor necrosis factor alpha, although the time course was considerably prolonged.

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).

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.

scholarly journals Characterization of human tracheal epithelial cells transformed by an origin-defective simian virus 40.

1988 ◽  
Vol 85 (16) ◽  
pp. 5951-5955 ◽  
Author(s):  
D. C. Gruenert ◽  
C. B. Basbaum ◽  
M. J. Welsh ◽  
M. Li ◽  
W. E. Finkbeiner ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Tracheal Epithelial Cells ◽  
Tracheal Epithelial

scholarly journals Simian virus 40-transformed adherent cells from human long-term marrow cultures: cloned cell lines produce cells with stromal and hematopoietic characteristics

Blood ◽  
1987 ◽  
Vol 70 (2) ◽  
pp. 464-474 ◽  
Author(s):  
JW Singer ◽  
P Charbord ◽  
A Keating ◽  
J Nemunaitis ◽  
G Raugi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Stromal Cells ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Adherent Cells ◽  
Semisolid Medium ◽  
Microscopic Appearance ◽  
Marrow Cultures

Abstract Adherent cells from long-term marrow cultures from 23 individuals were transformed with wild-type simian virus 40 (SV40). After transformation, cloned cell lines were developed that even after rigorous subcloning invariably produced both stromal cells and round cells. The stromal cells expressed cytoskeletal filaments similar to those of long-term marrow culture adherent cells and produced interstitial and basal lamina collagen types. The round cells had the electron microscopic appearance of primitive hematopoietic cells and when examined with cytochemical stains and monoclonal antibodies to hematopoietic differentiation antigens had reaction patterns suggestive of cells from several lineages. Most round cells expressed the pan- hematopoietic T-200 determinant, and lesser percentages expressed the early T cell antigens CD-1 and CD-3, HLA-DR determinants, the monocytic antigen recognized by Leu M3, and the myeloid antigens detected by monoclonal antibodies 1G10 and 12.8. In addition, when plated in semisolid medium in the presence of a source of colony-stimulating activity, up to 11% of the cells formed colonies consisting of blastlike cells that also expressed hematopoietic cell surface determinants. The data suggest that adherent cells in long-term marrow cultures contain a cell that after transformation by SV40 obligately produces cells with hematopoietic as well as stromalike features.

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