SURFACE ANTIGENS OF MAMMALIAN CELLS AS GENETIC MARKERS. II

A second surface antigen, BL, lethal in the presence of specific antibody and complement has been identified on some human cells and shown to behave as a good genetic marker. It is autosomal, unlinked to the human AL antigen previously described, and unlinked to 15 other human genes. The AL antigen, which is linked to the lactic dehydrogenase A gene, is found on the HeLa, the cultured human fibroblast, and in small amounts on the human lymphocyte. BL occurs on HeLa cells, on cultured human fibroblasts, and on human lymphocytes, but not on human RBCs. Hybrid cells formed by fusion of human and Chinese hamster cells have been prepared containing each of the four possible combinations of these two markers. Highly selective antisera sensitive to each marker separately can be obtained. The use of single-cell plating to demonstrate the presence of the antigens and of hybrid cells containing desired combinations of the markers facilitates study in this system.

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Transfer of human genes conferring resistance to methylating mutagens, but not to UV irradiation and cross-linking agents, into Chinese hamster ovary cells

Molecular and Cellular Biology ◽

10.1128/mcb.7.5.2024-2030.1987 ◽

1987 ◽

Vol 7 (5) ◽

pp. 2024-2030

Author(s):

B Kaina ◽

A A Van Zeeland ◽

C Backendorf ◽

H W Thielmann ◽

P Van de Putte

Keyword(s):

Chinese Hamster Ovary ◽

Chinese Hamster Ovary Cells ◽

Human Fibroblasts ◽

Chinese Hamster ◽

Native Form ◽

Ovary Cells ◽

Human Dna ◽

Human Genes ◽

Partial Inactivation ◽

High Doses

Chinese hamster ovary cells were transfected by human DNA ligated to the bacterial gpt (xanthine-guanine-phosphoribosyltransferase) gene which was used either in its native form or after partial inactivation with methylnitrosourea. The gpt+ transfectants were screened for resistance to high doses of N-methyl-N'-nitro-N-nitrosoguanidine. Using this approach, we showed that Chinese hamster ovary cells can acquire N-methyl-N'-nitro-N-nitrosoguanidine resistance upon transfection with DNA from diploid human fibroblasts, that this resistance is transferable by secondary transfection and is specific for methylating mutagens, and that it is not caused by increased removal of O6-methylguanine, 3-methyladenine, and 7-methylguanine from DNA.

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Genetics of somatic mammalian cells. XV. Evidence for linkage between human genes for lactic dehydrogenase B and serine hydroxymethylase

Journal of Cellular Physiology ◽

10.1002/jcp.1040800217 ◽

1972 ◽

Vol 80 (2) ◽

pp. 291-297 ◽

Cited By ~ 24

Author(s):

Carol Jones ◽

Paul Wuthier ◽

Fa-Ten Kao ◽

Theodore T. Puck

Keyword(s):

Mammalian Cells ◽

Lactic Dehydrogenase ◽

Human Genes

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Transfer of human genes conferring resistance to methylating mutagens, but not to UV irradiation and cross-linking agents, into Chinese hamster ovary cells.

Molecular and Cellular Biology ◽

10.1128/mcb.7.5.2024 ◽

1987 ◽

Vol 7 (5) ◽

pp. 2024-2030 ◽

Cited By ~ 16

Author(s):

B Kaina ◽

A A Van Zeeland ◽

C Backendorf ◽

H W Thielmann ◽

P Van de Putte

Keyword(s):

Chinese Hamster Ovary ◽

Chinese Hamster Ovary Cells ◽

Human Fibroblasts ◽

Chinese Hamster ◽

Native Form ◽

Ovary Cells ◽

Human Dna ◽

Human Genes ◽

Partial Inactivation ◽

High Doses

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Toxicity testing in vitro. I. The effects of Δ9-tetrahydrocannabinol and aflatoxin B1 on the growth of cultured human fibroblasts

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y76-075 ◽

1976 ◽

Vol 54 (4) ◽

pp. 541-545 ◽

Cited By ~ 2

Author(s):

James T. Cooper ◽

Samuel Goldstein

Keyword(s):

Human Fibroblasts ◽

Toxicity Testing ◽

Plating Efficiency ◽

Cultured Human Fibroblasts ◽

Mass Cultures ◽

Aflatoxin B ◽

Diploid Cells ◽

Dose Levels ◽

Cell Plating

The acute toxicity of Δ9-tetrahydrocannabinol (Δ9-THC) and aflatoxin B1 to two strains of cultured human fibroblasts has been studied. Δ9-THC had no effect on cell plating efficiency or on the growth of mass cultures at doses of 1 μg/ml (3.18 μM) or less; at 10 μg/ml plating efficiency was reduced by approximately half and at 20 μg/ml colony formation was zero. Aflatoxin B1 reduced plating efficiency at dose levels of 0.1 μg/ml (0.32 μM) and above; in mass cultures it retarded growth at 1 μg/ml and produced complete inhibition at 5 μg/ml. The potential usefulness of cultured human fibroblasts in toxicity testing is discussed and the importance of using normal diploid cells rather than aneuploid permanent lines is emphasized. The limitations of cell cultures in assessing toxicity, and possible solutions to these are considered.

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Regulation of the cell cycle by the cdk2 protein kinase in cultured human fibroblasts.

The Journal of Cell Biology ◽

10.1083/jcb.121.1.101 ◽

1993 ◽

Vol 121 (1) ◽

pp. 101-111 ◽

Cited By ~ 167

Author(s):

M Pagano ◽

R Pepperkok ◽

J Lukas ◽

V Baldin ◽

W Ansorge ◽

...

Keyword(s):

Cell Cycle ◽

Protein Kinase ◽

Mammalian Cells ◽

Human Fibroblasts ◽

S Phase ◽

Gene Products ◽

Related Gene ◽

Cultured Human Fibroblasts ◽

G2 Phase ◽

The Relationship

In mammalian cells inhibition of the cdc2 function results in arrest in the G2-phase of the cell cycle. Several cdc2-related gene products have been identified recently and it has been hypothesized that they control earlier cell cycle events. Here we have studied the relationship between activation of one of these cdc2 homologs, the cdk2 protein kinase, and the progression through the cell cycle in cultured human fibroblasts. We found that cdk2 was activated and specifically localized to the nucleus during S phase and G2. Microinjection of affinity-purified anti-cdk2 antibodies but not of affinity-purified anti-cdc2 antibodies, during G1, inhibited entry into S phase. The specificity of these effects was demonstrated by the fact that a plasmid-driven cdk2 overexpression counteracted the inhibition. These results demonstrate that the cdk2 protein kinase is involved in the activation of DNA synthesis.

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THE INTERACTION OF MAMMALIAN CELLS WITH ANTIBODIES. I

Journal of Experimental Medicine ◽

10.1084/jem.113.3.599 ◽

1961 ◽

Vol 113 (3) ◽

pp. 599-610 ◽

Cited By ~ 28

Author(s):

M. Oda ◽

T. T. Puck

Keyword(s):

Mammalian Cells ◽

Chinese Hamster ◽

Cell Killing ◽

Antibody Activity ◽

Specific Cell ◽

Plating Technique ◽

Killing Activity ◽

Killing Action ◽

Very High ◽

Cell Plating

The single cell plating technique has been applied to quantitation of the reproductive killing of mammalian cells by specific antibodies. This method confirms previous demonstrations by other workers of localization of all the killing activity in the γ-globulin fraction of specific cell antisera but not of normal sera; the need for complement for the killing action in low doses of antibody and the leakage of cell constituents attending cell killing under these conditions. In concentrations of 4 per cent or higher of heated antiserum cell killing occurs without added complement. The cell plating technique permits highly reproducible quantitation of antibody action and demonstrates antibody activity in sera diluted 1:3000. It permits demonstration of very high degrees of species specificity as shown by virtually complete absence of cross-reaction between antisera to Chinese hamster and S3 HeLa cells, respectively. Somatic cells which have been sensitized by absorption of specific antibody lose their sensitization when incubated at 37° unless complement is added within 1 hour.

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Ultrastructural Changes in Three Different Mammalian Cells Following Ultraviolet Laser Irradiation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100095017 ◽

1972 ◽

Vol 30 ◽

pp. 350-351

Author(s):

K. Shankar Narayan ◽

Kailash C. Gupta ◽

Tohru Okigaki

Keyword(s):

Ultraviolet Light ◽

Mammalian Cells ◽

Biological Effects ◽

Survival Rates ◽

Chinese Hamster ◽

Cell Types ◽

Differential Sensitivity ◽

Ultrastructural Changes ◽

Ultraviolet Laser

The biological effects of short-wave ultraviolet light has generally been described in terms of changes in cell growth or survival rates and production of chromosomal aberrations. Ultrastructural changes following exposure of cells to ultraviolet light, particularly at 265 nm, have not been reported.We have developed a means of irradiating populations of cells grown in vitro to a monochromatic ultraviolet laser beam at a wavelength of 265 nm based on the method of Johnson. The cell types studies were: i) WI-38, a human diploid fibroblast; ii) CMP, a human adenocarcinoma cell line; and iii) Don C-II, a Chinese hamster fibroblast cell strain. The cells were exposed either in situ or in suspension to the ultraviolet laser (UVL) beam. Irradiated cell populations were studied either "immediately" or following growth for 1-8 days after irradiation.Differential sensitivity, as measured by survival rates were observed in the three cell types studied. Pattern of ultrastructural changes were also different in the three cell types.

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