Establishment and characterization of tracheal epithelial cell lines, TM01 and TM02-3, from transgenic mice bearing temperature-sensitive simian virus 40 large T-antigen gene.
Murine tracheal epithelial cell lines, TM01 and TM02-3, were established from a primary culture of tracheal cells of adult transgenic mice bearing a temperature-sensitive simian virus (SV40) large T-antigen gene. Both TM01 and TM02-3 cells, which grew until confluent monolayers were formed, maintained tight contact with neighboring cells, and retained the characteristics of epithelial cells with microvilli on the surface. These cells grew at a permissive temperature (33 degrees C), but did not at a nonpermissive temperature (39 degrees C), indicating that TM01 and TM02-3 cells undergo temperature-sensitive growth. Large T-antigen was expressed only in the nuclei at 33 degrees C. Sepharose CL-4B column chromatography using a 14C-glucosamine hydrochloride, indicating that both cells produced high molecular weight glycoconjugates, and suggesting that these cells may originate from mucus-producing cells. TM01 cells expressed intercellular adhesion molecular-1 (ICAM-1) in both unstimulated and stimulated (1,000 U/ml tumor necrosis factor-alpha and 500 U/ml interferon-gamma) conditions, whereas TM02-3 cells expressed ICAM-1 only under stimulated conditions. We conclude that these cell lines may serve as a useful model to study the tracheal cell functions under defined in vitro conditions.
Sugiyama
N
N
Basic Technology Research Laboratory, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
Tabuchi
Y
Y
Numata
F
F
Uchida
Y
Y
Horiuchi
T
T
Ishibashi
K
K
Ono
S
S
Obinata
M
M
Furusawa
M
M
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D002250:Carbon Radioisotopes / Q000302:isolation & purification / Q000378:metabolism; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002847:Chromatography, Agarose; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D005944:Glucosamine / Q000302:isolation & purification / Q000378:metabolism; D006001:Glycoconjugates / Q000096:biosynthesis; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D013696:Temperature; D014132:Trachea / Q000737:chemistry / Q000166:cytology* / Q000254:growth & development* / Q000378:metabolism / Q000648:ultrastructure; D014410:Tumor Stem Cell Assay
D000952:Antigens, Polyomavirus Transforming; D002250:Carbon Radioisotopes; D006001:Glycoconjugates; D018799:Intercellular Adhesion Molecule-1; D005944:Glucosamine