A Study of the Penetration of Mammalian Cells by Deoxyribonucleic Acids

Borenfreund, Ellen; Bendich, Adrianne

doi:10.1083/jcb.9.1.81

Cited by 61 publications

(13 citation statements)

References 27 publications

(36 reference statements)

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“…If this difference is real, it could represent reutilization of labeled FL cell DNA, or FL cell D N A breakdown product. The phenomenon of D N A reutilization has been suggested in a n u m b e r of other systems (4-7, 9, 10, 16, 23), in which tritium-labeled DNA, available as purified D N A (4,16,23) or available in nuclei of infused leukocytes (5,7) or in degenerating liver cells or lymphocytes (6,23), appeared in the nuclei of rapidly dividing transplanted tumor cells, or in dividing cells of the gut, testis, etc. An obvious parallel to these observations exists in our own experiments in which degenerating FL cells are typically present in the center of FL colonies (2), and could serve as a source of diffusible label to multiplying fibroblasts.…”

Section: Bone Induction By Labeled Fl Cellsmentioning

confidence: 95%

Bone Formation Induced in Mouse Thigh by Cultured Human Cells

Anderson

Coulter

1967

The Journal of Cell Biology

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Cultured FL human amnion cells injected intramuscularly into cortisone-conditioned mice proliferate to form discrete nodules which become surrounded by fibroblasts. Within 12 days, fibroblastic zones differentiate into cartilage which calcifies to form bone. Experiments were conducted to test the hypothesis that FL cells behave as an inductor of bone formation. In the electron microscope, FL cells were readily distinguished from surrounding fibroblasts. Transitional forms between the two cell types were not recognized. Stains for acid mucopolysaccharides emphasized the sharp boundary between metachromatic fibroblastic and cartilaginous zones and nonmetachromatic FL cells. 85S was taken up preferentially by fibrob!asts and chondrocytes and then deposited extracellularly in a manner suggesting active Secretion of sulfated mucopolysaccharides. FL cells showed negligible 35S utilization and secretion. FL cells, labeled in vitro with thymidine-aH, were injected and followed radioautographically, during bone formation. Nuclear label of injected FL cells did not appear in adjacent fibroblasts in quantities sufficient to indicate origin of the latter from FL cells. The minimal fibroblast nuclear labeling seen may represent reutilization of label from necrotic FL cells. It is suggested that FL cells injected into the mouse thigh induced cartilage and bone formation by host fibroblasts.

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Section: Bone Induction By Labeled Fl Cellsmentioning

confidence: 95%

Bone Formation Induced in Mouse Thigh by Cultured Human Cells

Anderson

Coulter

1967

The Journal of Cell Biology

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“…Penetratiorvof mammalian cells by DNA has been observed in a number of laboratories. [13][14][15][16][17][18] Reports have also appeared concerning the physiological effects of exogenous DNA. Three different groups have observed genetic transformation of mammalian cells by mammalian DNA in tissue culture.5,19,20 However, the cells transformed represented only a tiny fraction of the total number exposed to the DNA.…”

Section: Discussionmentioning

confidence: 99%

SECTION OF BIOLOGICAL AND CHEMICAL SCIENCES: THE ACTION OF THYMUS DNA ON L1210 LEUKEMIA CELLS*,†

Glick

Goldberg

1966

Trans NY Acad Sci

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We have recently reported the inhibition of L1210 tumor growth in mice by preincubating the L1210 cells with mouse thymus DNA.l Treating the DNA with DNase abolished its growth inhibitory effect. Intact L1210 DNA was also ineffective. This work and additional ~t u d i e s~'~ have indicated that the base sequence, helical structure, and size of the thymus DNA a r e all important factors in causing inhibitionof tumor growth. Our current experiments deal with the action of DNA on L1210 cells in vitro. MATERIALS AND METHODSMouse thymus DNA was isolated and treated with RNase by the method of Szybalska and S~y b a l s k i .~ DNA was similarly prepared from L1210 leukemia cells. Except where indicated, the L1210 cells were grown in spinner cultures containing medium RPMI 1630 + 5 per cent fetal calf serum.6 (This medium includes 20 amino acids, a number of vitamins, but no purine or pyrimidine derivatives.) In most cases the cells were washed and incubated at 37°C in 4 ml. of a phosphate-buffered saline (pH 7.5)5 containing 5.5 mM glucose and 0.2 mM spermine tetrahydrochloride. The cells were usually treated for 30 minutes with DNA (30 to60 pgper lo6 cells) and then washed and resuspended in DNA-free PBSG (phosphate-buffered saline -t glucose). Control cells were incubated for 30 minutes without DNA and then washed and resuspended in fresh PBSG. Between two and 20 x lo6 cells per ml. were used for experiments in vivo. About 106 cells per ml. were used for the experiments in vitro, with the exception of the isotope-incorporation experiments in which 2-4 x lO'.cells per ml. were utilized.Cell viability was determined by adding 0.2 ml. of 0.1 per cent trypan blue (dissolved in Hanks Balanced Salt Solution) to 0.2 ml. of cell suspension. Only unstained cells were considered ~i a b l e .~,~ Cells were counted microscopically with a Speirs-Levy Chamber and electronically by means of a Coulter Counter.

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“…The indication that intact DNA was incorporated in some experiments gained support from the studies of Borenfreund and Bendich (19) who employed an antibody against DNase to demonstrate the incorporation of polymerized DNA by HeLa cells in vitro. Kay (20) has reported that once DNA is incorporated it may be utilized without intracellular degradation.…”

Section: Conditions and Significance Of Reutilization Of Dna And Its mentioning

confidence: 91%

THE IN VIVO REUTILIZATION OF LYMPHOCYTIC AND SARCOMA DNA BY CELLS GROWING IN THE PERITONEAL CAVITY

Rieke

1962

The Journal of Cell Biology

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An ascites tumor, Sarcoma I, was transplanted to isologous and homologous mice which had been labeled with tritiated thymidine from 1 to 24 hours previously. Radioautographic preparations revealed labeled host lymphocytes emerging to mingle with the transplanted tumor and the subsequent appearance of nuclear radioactivity in the sarcoma. Sarcoma cells cultured subcutaneously or in Millipore diffusion chambers in previously labeled mice did not demonstrate significant radioactivity. Transplantation of washed, Ha-thymidinclabeled lymphocytes to non-radioactive, sarcoma-bearing mice was followed by the gradual appearance of nuclear radioactivity in the sarcoma. The label in the sarcoma was entirely removed by deoxyribonuclcase but not by ribonuclease treatment prior to radioautography. Intraperitoneal injections of purified, Ha-thymidine-labeled sarcoma or lymphoid DNA in normal or tumor-bearing mice were followed by radioactivity appearing in sarcoma or normal peritoneal mononuclear cells. It was concluded that rcutilization of DNA and its metabolites may occur in vivo, and the conditions under which reutilization may be detected are discussed.

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A Study of the Penetration of Mammalian Cells by Deoxyribonucleic Acids

Cited by 61 publications

References 27 publications

Bone Formation Induced in Mouse Thigh by Cultured Human Cells

Bone Formation Induced in Mouse Thigh by Cultured Human Cells

SECTION OF BIOLOGICAL AND CHEMICAL SCIENCES: THE ACTION OF THYMUS DNA ON L1210 LEUKEMIA CELLS*,†

THE IN VIVO REUTILIZATION OF LYMPHOCYTIC AND SARCOMA DNA BY CELLS GROWING IN THE PERITONEAL CAVITY

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