Effect of Daunorubicin and Adriamycin on Nucleic ACID Synthesis of Serum Stimulated Mouse Embryo Fibroblasts

1977 ◽  
Vol 63 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Rosanna Supino ◽  
Anna M. Casazza ◽  
Aurelio Di Marco
Keyword(s):  
Dna Synthesis ◽  
Mouse Embryo ◽  
Rna Synthesis ◽  
Calf Serum ◽  
Acid Synthesis ◽  
Anthracycline Antibiotics ◽  
Simple Method ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts

This paper reports the effects of daunorubicin and adriamycin on DNA and RNA synthesis of in vitro cultured mouse embryo fibroblasts (MEF) stimulated by fetal calf serum (FCS). The addition of FCS to quiescent MEF cultures brings about a wave of RNA synthesis, followed by DNA synthesis which starts between 8 and 12 h after change of medium and proceed for up to 24 h. These cells are therefore partially synchronized. The level of DNA synthesis depends on the amount of FCS added. Daunorubicin and adriamycin are almost equally effective in inhibiting DNA synthesis, as well as cell proliferation, which takes place later. Adriamycin is more active than daunorubicin on RNA synthesis. In cultures treated for an 8 h period starting at different times after FCS addition, the highest DNA synthesis inhibition is achieved by treatment during the first 8 h, when DNA synthesis has not yet started. The cellular uptake of daunorubicin is constantly higher than that of adriamycin, in any experimental condition tested. The results show that FCS-stimulated MEF can provide a simple method for studying the effects of anthracycline antibiotics on partially synchronized cells.

scholarly journals Continuous replication of Friend virus complex (spleen focus-forming virus-lymphatic leukemia-inducing virus) in mouse embryo fibroblasts. Retention of leukemogenicity and loss of immunosuppressive properties.

1975 ◽  
Vol 142 (4) ◽  
pp. 936-948 ◽  
Author(s):  
R J Eckner
Keyword(s):  
Mouse Embryo ◽  
Helper Virus ◽  
Productive Infection ◽  
Friend Virus ◽  
Focus Formation ◽  
Lymphatic Leukemia ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts

Exposure of NIH Swiss mouse embryo fibroblasts (MEF) to infectious Friend virus (FV) complex [containing defective spleen focus-forming virus (SFFV) and endogenous NB-tropic leukemia-inducing helper virus (LLV-F)] resulted in the productive infection of these cells by both SFFV and LLV-F. Stocks of SFFV derived after extensive growth in this Swiss MEF cell culture system are fully leukemogenic in adult mice for the induction of erythroleukemia and spleen foci. In addition, in vitro-derived LLV-F, when isolated free of SFFV, is fully leukemogenic for the induction of lymphatic leukemia when inoculated into susceptible newborn BALB/c mice. The host range of in vitro-derived FV complex (i.e., FV-TC) for focus formation in vivo is NB-tropic. Unlike in vivo-derived FV complex, FV-TC does not suppress the responsiveness of murine thymocytes to concanavalin A (Con A) in vitro. Rather, FV-TC acts as a mitogen to nonspecifically stimulate the proliferation of BALB/c thymocytes. The mitogenicity of in vitro-derived FV complex is directly associated with the presence of type-C virus particles, is a heat-labile and UV-sensitive property of the virus, and may be primarily due to LLV since equivalent amounts of LLV with or without SFFV present are equally mitogenic. One in vivo passage of FV-TC resulted in the total loss of this mitogenic property with the reappearance of full immunosuppressive properties. This result demonstrates a clear association between in vivo growth of FV and its ability to suppress mouse thymocytes, and suggests that FV complex (SFFV-LLV) is not inherently immunosuppressive for these cells. While the mechanism of this interconversion between immunostimulatory and fully suppressive virus is unknown, both virus markers appear to be dependent upon the presence of infectious FV.

scholarly journals EFFECT OF GROWTH HORMONE ON THE METABOLISM OF THYMUS AND ON THE IMMUNE RESPONSE AGAINST SHEEP ERYTHROCYTES

1971 ◽  
Vol 134 (5) ◽  
pp. 1095-1113 ◽  
Author(s):  
M. R. Pandian ◽  
G. P. Talwar
Keyword(s):  
Growth Hormone ◽  
Immune Response ◽  
Dna Synthesis ◽  
Rna Synthesis ◽  
Calf Serum ◽  
Lymphoid Organs ◽  
Stimulatory Action ◽  
Early Action ◽  
Stimulation Of

The effect of pituitary growth hormone on the biosynthesis of DNA in the thymus and other lymphoid organs, as well as the ability of the rat to respond immunologically to sheep red blood cells, has been evaluated. There is a marked reduction in plaque-forming cells, hemagglutination titers, and DNA synthesis in animals when examined at 15 wk after hypophysectomy. Administration of bovine growth hormone (BGH) leads to the enhancement of DNA synthesis in lymphoid organs and recovery of the immune response. Similar effects of the hormone are observed in plateaued rats. Injection of rabbit anti-BGH globulins, in contrast to normal rabbit globulins, over 5 days causes a drop in the weight of the thymus and in the rate of DNA synthesis in this organ. The thymus is also the organ in which stimulation of DNA synthesis is observed at a time period earlier than the spleen and lymph nodes after a single injection of BGH. The hormone stimulates not only the incorporation of thymidine-3H into DNA in the cortical cells, but also the incorporation of sodium sulfate-35S into TCA-insoluble biopolymers reported to be elaborated in the medullary area of the thymus. An in vitro system for the action of BGH on the thymus has been described. There is an obligatory requirement for calcium, but not for fetal calf serum in the medium for the hormone effect. An early action of the hormone is the enhanced incorporation of uridine-G-3H into RNA in thymocytes which is followed by a stimulation of the synthesis of proteins and DNA. The stimulatory action of growth hormone on RNA synthesis is not because of a facilitated uptake of the radioactive uridine by the cells under hormonal influence, a mechanism by which insulin is observed to increase RNA synthesis in thymocytes in vitro. The action of growth hormone on thymocytes is specific, since thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and heat-inactivated growth hormone are not effective. BGH has also a beneficial action on the regeneration of the thymus and spleen in starved rats.

The reproductive potential of normal mouse embryo fibroblasts during culture in vitro

1984 ◽  
Vol 66 (1) ◽  
pp. 401-409
Author(s):  
C. Karatza ◽  
S. Shall
Keyword(s):  
Mouse Embryo ◽  
Colony Size ◽  
Reproductive Potential ◽  
Cell Strain ◽  
Growth Fraction ◽  
Growth Behaviour ◽  
Direct Estimate ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts

A direct estimate of the reproductive potential of mouse embryo fibroblasts through their entire lifespan has been made using the mini-clone technique, which permits the direct observation of the growth fraction in a bulk population by inspection of the growth behaviour of individual cells. We have measured the colony size on each island that contained one or two cells at the beginning and the fraction of islands which, starting from one or two cells failed to divide even once. We observed that even in a young culture there are individual cells that can only reproduce two or three times. With each succeeding passage the distribution of colony sizes shifts to a greater proportion of small colonies. The median colony size decreases with each passage. Furthermore, the fraction of non-dividers directly observed increases smoothly with time; the fraction of non-dividers is quite small at the first passage but increases steadily to reach 0.6 at the last passage, after about 30 generations. These smooth changes in the growth behaviour of this cell strain are accurately described by the mortalization theory of Shall and Stein, in which the single parameter gamma, describes the change in reproductive potential over the entire lifespan. The parameter gamma describes the rate at which the doubling time of the culture increases; it is the number of generations at which half of the newborn cells are themselves reproductively sterile. Our present data provide an estimate of gamma for this cell strain equal to 21.2 generations, which compares well with a previous estimate of 20.3 generations.

Effect of Syngeneic and Allogeneic Lymphocytes on L-cells and Mouse Embryo Fibroblasts in vitro

Nature ◽  
1967 ◽  
Vol 214 (5094) ◽  
pp. 1229-1230 ◽  
Author(s):  
INESSA YU. CHERNYAKHOVSKAYA ◽  
ZAIRA G. KADAGGIDZE ◽  
GEORGE J. SVET-MOLDAVSKY
Keyword(s):  
Mouse Embryo ◽  
L Cells ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts ◽  
Allogeneic Lymphocytes

scholarly journals 5′-AMP-Activated Protein Kinase (AMPK) Is Induced by Low-Oxygen and Glucose Deprivation Conditions Found in Solid-Tumor Microenvironments

2006 ◽  
Vol 26 (14) ◽  
pp. 5336-5347 ◽  
Author(s):  
Keith R. Laderoute ◽  
Khalid Amin ◽  
Joy M. Calaoagan ◽  
Merrill Knapp ◽  
Theresamai Le ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mouse Embryo ◽  
Cellular Response ◽  
Null Mouse ◽  
Low Oxygen ◽  
Tumor Microenvironments ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts ◽  
Amp Activated Protein Kinase

ABSTRACT Low oxygen gradients (hypoxia and anoxia) are important determinants of pathological conditions under which the tissue blood supply is deficient or defective, such as in solid tumors. We have been investigating the relationship between the activation of hypoxia-inducible factor 1 (HIF-1), the primary transcriptional regulator of the mammalian response to hypoxia, and 5′-AMP-activated protein kinase (AMPK), another regulatory system important for controlling cellular energy metabolism. In the present study, we used mouse embryo fibroblasts nullizygous for HIF-1α or AMPK expression to show that AMPK is rapidly activated in vitro by both physiological and pathophysiological low-oxygen conditions, independently of HIF-1 activity. These findings imply that HIF-1 and AMPK are components of a concerted cellular response to maintain energy homeostasis in low-oxygen or ischemic-tissue microenvironments. Finally, we used transformed derivatives of wild-type and HIF-1α- or AMPKα-null mouse embryo fibroblasts to determine whether AMPK is activated in vivo. We obtained evidence that AMPK is activated in authentic hypoxic tumor microenvironments and that this activity overlaps with regions of hypoxia detected by a chemical probe. We also showed that AMPK is important for the growth of this tumor model.

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