scholarly journals ACTION OF X-RAYS ON MAMMALIAN CELLS

1957 ◽  
Vol 106 (4) ◽  
pp. 485-500 ◽  
Author(s):  
Theodore T. Puck ◽  
Dimitry Morkovin ◽  
Philip I. Marcus ◽  
Steven J. Cieciura

Survival curves of normal human cells from a variety of tissues exposed to varying doses of x-irradiation have been constructed, which permit definition of the intrinsic radiation sensitivity of the reproductive power of each cell type. The mean lethal dose of x-irradiation for all the cells employed, including those from normal and cancerous organs, those exhibiting diploid and polyploid chromosome number; those from embryonic and adult tissues, including recently isolated cells and cultures which had been maintained in vitro for many years, and cells exhibiting either epithelioid or fibroblastic morphology, was found to be contained between the limits of 50 to 150 r. Other similarities in the pattern of radiation effects, such as giant formation and abortive colonial growth, in these cells and that of the HeLa S3, previously studied, confirm the hypothesis that the pattern of reaction to x-irradiation previously elucidated, is representatative, at least in over-all outline, for a large variety of human cells. While the radiation survival curves of various human cells are similar in the gross, small but important characterizing differences have been found. All epithelioid cells so far studied are approximately 2-hit, and more radioresistant than the fibroblast-like cells whose survival data correspond to a mean lethal dose of around 60 r, and which so far can be fitted by either 1-hit or 2-hit curves. The earlier prediction that the major radiobiologic damage to mammalian cells is lodged in the genetic apparatus was confirmed by the demonstration of high frequency of mutants among the survivors of doses of 500 to 900 r. All the data on the x-radiosensitivity of these cells can be explained on the basis of a defect resulting from primary damage localized in one or more chromosomes. These considerations afford a convincing explanation of several aspects of the mammalian radiation syndrome.

1985 ◽  
Vol 8 (6) ◽  
pp. 517-522 ◽  
Author(s):  
T. J. FitzGerald ◽  
C. Daugherty ◽  
K. Kase ◽  
L. A. Rothstein ◽  
M. McKenna ◽  
...  

1961 ◽  
Vol 200 (5) ◽  
pp. 1039-1042 ◽  
Author(s):  
Bernard D. Newsom ◽  
Donald J. Kimeldorf

A lethal dose of X irradiation can be made sublethal by the fractionation of a dose into a series of smaller doses separated by adequate time intervals. The results reported here demonstrate how an environmental stress, such as cold, may affect this mitigating action of dose fractionation. Rats were maintained at 6 C while receiving a total dose of 600 r in one or eight exposures with various intervals between exposures. A greater interval was required between radiation exposures at 6 C than at 23 C to reduce the lethal response following irradiation. From the results of the food consumption measurements, it appears that cold exposure emphasizes the consequence of the reduced food intake which follows daily doses of 75 r in the rat. The irradiated rat was able to increase the amount of food consumed during cold exposure; however, the increase was not sufficient for adaptation. The lowered consumption which followed eight daily 75-r exposures was sufficient to cause death in pair-fed rats maintained at 6 C, but not at 23 C.


Nature ◽  
1973 ◽  
Vol 243 (5408) ◽  
pp. 450-453 ◽  
Author(s):  
CARMIA BOREK ◽  
ERIC J. HALL
Keyword(s):  

1956 ◽  
Vol 103 (2) ◽  
pp. 273-284 ◽  
Author(s):  
Theodore T. Puck ◽  
Philip I. Marcus ◽  
Steven J. Cieciura

Two methods for simple and rapid plating of single HeLa cells, human, carcinomatous cells, are described. These result in growth and formation of colonies from each single cell. One of these procedures uses irradiated, non-multiplying "feeder" cells to condition the medium. The second requires more gentle handling of the cells, but otherwise is virtually the same as that used in plating bacteria on semisolid, nutrient media. By extension of these methods, it is possible to isolate single mutant colonies and grow pure clonal stocks of animal cells. These genetically uniform strains are much more homogeneous in their behavior than the parental HeLa cell population. Growth curves obtained from developing colonies are highly reproducible. The most active mutant stocks so far isolated display a generation time of 18 to 20 hours. In pooled human serum HeLa cells assume a highly stretched, ameboid form, with marked motility; whereas growth of the same cells in a variety of non-human sera results in tightly packed, columnar, epithelial-like morphology. The two cell types possess volumes, nuclear cross-sections, plating efficiencies, and generation times which are identical within experimental error, but display widely different cross-sectional areas, suggesting that the basic change occurs in the cell surface. It is conceivable that this change may be related to that which enables the cells of a compact tumor to become invasive. Animal cells subjected to the standard trypsinization procedures which involve mechanical trauma and repeated washings in incomplete media leak large amounts of P and suffer impaired ability to reproduce as isolated cells. Application of the methods described in this paper as a tool for quantitative study of normal mammalian cell growth, physiology, genetics, and biochemistry, and the response of cells to drugs, viruses, high energy radiation, and other agents have been indicated.


2002 ◽  
Vol 184 (13) ◽  
pp. 3657-3663 ◽  
Author(s):  
Stephanie M. Stoll ◽  
Daniel S. Ginsburg ◽  
Michele P. Calos

ABSTRACT We demonstrate that the site-specific integrase encoded by phage TP901-1 of Lactococcus lactis subsp. cremoris has potential as a tool for engineering mammalian genomes. We constructed vectors that express this integrase in Escherichia coli and in mammalian cells and developed a simple plasmid assay to measure the frequency of intramolecular integration mediated by the integrase. We used the assay to document that the integrase functions efficiently in E. coli and determined that for complete reaction in E. coli, the minimal sizes of attB and attP are 31 and 50 bp, respectively. We carried out partial purification of TP901-1 integrase protein and demonstrated its functional activity in vitro in the absence of added cofactors, characterizing the time course and temperature optimum of the reaction. Finally, we showed that when expressed in human cells, the TP901-1 integrase carries out efficient intramolecular integration on a transfected plasmid substrate in the human cell environment. The TP901-1 phage integrase thus represents a new reagent for manipulating DNA in living mammalian cells.


Sign in / Sign up

Export Citation Format

Share Document