TY - JOUR
T1 - Mouse cell clones for improved quantitation of carcinogen-induced altered differentiation
AU - Kulesz-martin, Molly
AU - Yoshida, Mitsuaki A.
AU - Prestine, Laura
AU - Yuspa, Stuart H.
AU - Bertram, John S.
N1 - Funding Information:
The authors are grateful to Dr Henry Hennings of the National Cancer Institute for atomic absorption spectroscopy to determine calcium levels in culture medium, Dr Bonnie Asch, the Department of Experimental Pathology, RPMI, for performance of the indirect immunofluorescence tests for keratin, Dr Tung-Tien Sun, Department of Dermatology, NYU School of Medicine, for anti-keratin sera, Dr Avcry A.Sandberg, Chief, Genetics and Endocrinology, RPMI, for review of the chromosome studies, and Mae Brown for secretarial assistance. This investigation was supported by PHS grant Number RO1-CA-31101, and Core Grant CA-24538 awarded by the National Cancer Institute, DHHS.
PY - 1985/9
Y1 - 1985/9
N2 - Two new mouse epidermal cell lines have been isolated and characterized as target cells for three chemical carcinogens. The ability to grow these cells at low density ( ∼5 clonogenic cells/cm2) has permitted more precise quantitation of chemical carcinogen-induced changes in epidermal differentiation. The cell lines, designated 291 and 271c, retain the property previously observed in primary cultures of mouse epidermal cells, that is the regulation of terminal differentiation by extracellular Ca2+ ion. Altered response to extracellular Ca2+ after carcinogen treatment of these cells is the basis of the assay endpoint. Other normal properties demonstrated by these cells are keratin immunofluorescence patterns, ability to form cornified envelopes in response to Ca2+ and a lack of tumorigenicity. Both of the lines have high cloning efficiencies (up to 20%) and characteristic epidermal morphology. Their chromosome number, however, is near tetraploid. Dose response studies indicated an increase in colonies with altered response to Ca2+ proportional to the dose of three chemical carcinogens: DMBA 0.001 - 0.5 μg/ml × 24 h, MCA 0.01 - 5 μg/ml × 24 h and MNNG 0.01 - 0.2 μg/ml × 1 h. The optimized assay protocol has provided a reproducible means of quantitating carcinogen-altered epidermal cells relative to carcinogen dose, and of isolating cell clones for studies of altered differentiation in carcinogenesis and chemotherapy.
AB - Two new mouse epidermal cell lines have been isolated and characterized as target cells for three chemical carcinogens. The ability to grow these cells at low density ( ∼5 clonogenic cells/cm2) has permitted more precise quantitation of chemical carcinogen-induced changes in epidermal differentiation. The cell lines, designated 291 and 271c, retain the property previously observed in primary cultures of mouse epidermal cells, that is the regulation of terminal differentiation by extracellular Ca2+ ion. Altered response to extracellular Ca2+ after carcinogen treatment of these cells is the basis of the assay endpoint. Other normal properties demonstrated by these cells are keratin immunofluorescence patterns, ability to form cornified envelopes in response to Ca2+ and a lack of tumorigenicity. Both of the lines have high cloning efficiencies (up to 20%) and characteristic epidermal morphology. Their chromosome number, however, is near tetraploid. Dose response studies indicated an increase in colonies with altered response to Ca2+ proportional to the dose of three chemical carcinogens: DMBA 0.001 - 0.5 μg/ml × 24 h, MCA 0.01 - 5 μg/ml × 24 h and MNNG 0.01 - 0.2 μg/ml × 1 h. The optimized assay protocol has provided a reproducible means of quantitating carcinogen-altered epidermal cells relative to carcinogen dose, and of isolating cell clones for studies of altered differentiation in carcinogenesis and chemotherapy.
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U2 - 10.1093/carcin/6.9.1245
DO - 10.1093/carcin/6.9.1245
M3 - Article
C2 - 2411440
AN - SCOPUS:0022255225
SN - 0143-3334
VL - 6
SP - 1245
EP - 1254
JO - Carcinogenesis
JF - Carcinogenesis
IS - 9
ER -