scholarly journals STUDIES ON BONE MARROW IN VITRO

Blood ◽  
1948 ◽  
Vol 3 (2) ◽  
pp. 165-174 ◽  
Author(s):  
A. ROSIN ◽  
M. RACHMILEWITZ
Keyword(s):  
Bone Marrow ◽  
Oxygen Tension ◽  
Gas Mixtures ◽  
Cent Oxygen ◽  
Low Oxygen ◽  
Bone Marrow Cultures ◽  
Oxygen Tensions ◽  
Marrow Cultures

Abstract The effect of various oxygen tensions on explanted bone marrow fragments was studied. It was found that gas mixtures containing 1, 3, 5, 10 and 12 per cent oxygen have an injurious effect on hemic cells. Bone marrow maintained in these gas mixtures showed various degrees of degeneration, which was the more pronounced the lower the oxygen tension. Mitotic activity was also found to be reduced under the influence of low oxygen tension. Bone marrow cultures maintained in a gas mixture containing 15 per cent oxygen did not show appreciable changes and were similar to the controls. Increased rate of maturation and multiplication occurred in bone marrow cultures maintained in an excess of oxygen, i.e. 50 per cent. The significance of these findings in the light of observations on the effect of anoxia in vivo has been discussed, and reported findings on the effect of low oxygen tensions on other tissues in vitro have been briefly reviewed.

Thrombopoietin-Induced Stimulation of Megakaryocyte- Enriched Bone Marrow Cultures

1979 ◽  
Author(s):  
K. L. Kellar ◽  
B. L. Evatt ◽  
C. R. McGrath ◽  
R. B. Ramsey
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Bone Marrow Cells ◽  
Rabbit Plasma ◽  
Bone Marrow Cultures ◽  
Plasma Fractions ◽  
Marrow Cultures ◽  
Stimulation Of

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.

The detection of in vivo hematotoxicity of benzene by in vitro liquid bone marrow cultures

1981 ◽  
Vol 60 (2) ◽  
pp. 346-353 ◽  
Author(s):  
Kenichi Harigaya ◽  
Marilyn E. Miller ◽  
Eugene P. Cronkite ◽  
Robert T. Drew
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

scholarly journals Multistep virus-induced leukemogenesis in vitro: description of a model specifying three steps within the myeloblastic malignant process.

1984 ◽  
Vol 4 (1) ◽  
pp. 216-220 ◽  
Author(s):  
J M Heard ◽  
S Fichelson ◽  
B Sola ◽  
M A Martial ◽  
B Varet ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Leukemia Virus ◽  
Latency Period ◽  
In Vitro Production ◽  
Long Latency ◽  
Permanent Cell ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

A helper-independent Friend leukemia virus was used to infect bone marrow cultures. This virus induces myeloblastic leukemia in mice after a long latency period. Infection of the bone marrow cultures resulted in the in vitro production of myeloblastic leukemogenesis after a long latency period. Three steps were observed in the evolution of the infected cultures, and permanent cell lines were derived at each step. This allowed us to individualize three successive events in the course of the myeloblastic transformation: (i) an abnormal responsiveness to the physiological hormone granulo-macrophagic colony-stimulating factor, (ii) the acquisition of growth autonomy, and (iii) the acquisition of in vivo tumorigenicity.

In Vitro Studies of Erythropoiesis

Blood ◽  
1955 ◽  
Vol 10 (6) ◽  
pp. 612-615 ◽  
Author(s):  
E. D. THOMAS
Keyword(s):  
Bone Marrow ◽  
Oxygen Tension ◽  
In Vitro Studies ◽  
Cent Oxygen ◽  
In Vitro Synthesis ◽  
Heme Synthesis ◽  
Rabbit Bone ◽  
Oxygen Tensions ◽  
Effect Of Oxygen

Abstract The effect of various oxygen tensions on the in vitro synthesis of heme by rabbit bone marrow was measured. At levels above 4 per cent oxygen there was no effect of oxygen tension on heme synthesis. Total anoxia stopped heme synthesis completely. No level of oxygen tension was found to stimulate heme synthesis.

Multistep virus-induced leukemogenesis in vitro: description of a model specifying three steps within the myeloblastic malignant process

1984 ◽  
Vol 4 (1) ◽  
pp. 216-220
Author(s):  
J M Heard ◽  
S Fichelson ◽  
B Sola ◽  
M A Martial ◽  
B Varet ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Leukemia Virus ◽  
Latency Period ◽  
In Vitro Production ◽  
Long Latency ◽  
Permanent Cell ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

A helper-independent Friend leukemia virus was used to infect bone marrow cultures. This virus induces myeloblastic leukemia in mice after a long latency period. Infection of the bone marrow cultures resulted in the in vitro production of myeloblastic leukemogenesis after a long latency period. Three steps were observed in the evolution of the infected cultures, and permanent cell lines were derived at each step. This allowed us to individualize three successive events in the course of the myeloblastic transformation: (i) an abnormal responsiveness to the physiological hormone granulo-macrophagic colony-stimulating factor, (ii) the acquisition of growth autonomy, and (iii) the acquisition of in vivo tumorigenicity.

scholarly journals Cellular Activities in Tissue Cultures of Leukemic Human Bone Marrow

Blood ◽  
1964 ◽  
Vol 24 (4) ◽  
pp. 389-401 ◽  
Author(s):  
JOSEPH G. SINKOVICS ◽  
CLIFTON D. HOWE ◽  
C. C. SHULLENBERGER
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Cell ◽  
Giant Cells ◽  
Target Cells ◽  
Tissue Cultures ◽  
Autoimmune Attack ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Abstract 1) The growth from human leukemic bone marrow particles begins with the emigration of cells having the morphologic characteristics of erythrocytic, granulocytic, monocytic and lymphocytic elements. Proliferation of fibroblastic-like cells replaces these elements within 3 to 4 weeks. Various types of immature leukocytes occur in larger numbers, and persist longer, in tissue cultures of human leukemic bone marrow than in bone marrow cultures from patients with no hematological disease. 2) Lymphocytes from either leukemic or non-leukemic bone marrow cultures often appear to enter the cytoplasm of fibroblast-like cells, but those derived from cultures of chronic lymphocytic leukemic bone marrow appeared intracellularly in much larger numbers and persisted there longer. One of the mechanisms of this phenomenon may be the feeder layer principle. Extensive injury of certain fibroblast-like cells in relation to relatively few small lymphocytes suggests the possibility of an autoimmune attack. It is possible that both the lymphocytes and the "target cells" may disintegrate. 3) Two types of multinucleated giant cells have been observed. One type is syncytial in structure, suggesting a viral mechanism of initiation. Another type resembles the polyploid giant cells which are commonly seen in old tissue cultures of whatever origin, and in various malignant diseases in vivo as well as in vitro. 4) At times, poorly differentiated round cells may persist in old fibroblastic cultures. These cells may be derived from the original explant which continued to produce them, or they may originate from fibroblast-like cells reconverted into a morphologically undifferentiated mesenchymal cell type.

scholarly journals In vivo analysis of DNA-protein interactions on the human erythropoietin enhancer.

1997 ◽  
Vol 17 (2) ◽  
pp. 851-856 ◽  
Author(s):  
B Hu ◽  
E Wright ◽  
L Campbell ◽  
K L Blanchard
Keyword(s):  
Oxygen Tension ◽  
Protein Interactions ◽  
Proximal Promoter ◽  
Low Oxygen ◽  
Initiation Rate ◽  
Cobalt Exposure ◽  
In Cells ◽  
Dna Protein Interactions

The erythropoietin (EPO) gene is one of the best examples of a mammalian gene controlled by oxygen tension. The DNA elements responsible for hypoxia-induced transcription consist of a short region of the proximal promoter and a <50-bp 3' enhancer. The elements act cooperatively to increase the transcriptional initiation rate approximately 100-fold in response to low oxygen tension in Hep3B cells. Two distinct types of transactivating proteins have been demonstrated to bind the response elements in the human EPO enhancer in vitro: one shows hypoxia-inducible DNA binding activity, while the other activity binds DNA under normoxic and hypoxic conditions. We have investigated the DNA-protein interactions on the human EPO enhancer in living tissue culture cells that produce EPO in a regulated fashion (Hep3B) and in cells that do not express EPO under any conditions tested (HeLa). We have identified in vivo DNA-protein interactions on the control elements in the human EPO enhancer by ligation-mediated PCR technology. We show that the putative protein binding sites in the EPO enhancer are occupied in vivo under conditions of normoxia, hypoxia, and cobalt exposure in EPO-producing cells. These sites are not occupied in cells that do not produce EPO. We also provide evidence for a conformational change in the topography of the EPO enhancer in response to hypoxia and cobalt exposure.

scholarly journals Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures.

1989 ◽  
Vol 9 (9) ◽  
pp. 3973-3981 ◽  
Author(s):  
G V Borzillo ◽  
C J Sherr
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Chain Gene ◽  
Mouse Bone Marrow ◽  
Feeder Layers ◽  
Bone Marrow Cultures ◽  
B Lymphoid ◽  
Marrow Cultures

Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B-lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage.

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