T. R. Bradley

Lexington VA Health Care System

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G. S. Hodgson

MRC Laboratory of Molecular Biology

Ivan Bertoncello

University of Melbourne

A. B. Kriegler

Peter MacCallum Cancer Centre

D Metcalf

Walter and Eliza Hall Institute of Medical Research

William A. Robinson

University of Colorado Anschutz Medical Campus

Margaret A. Sumner

University of Melbourne

Ian A. Cooper

Defence Science and Technology Laboratory

Robert A. Roosa

California University of Pennsylvania

Phyllis Fry

University of Melbourne

Martin Rosendaal

Semmelweis University

Cooperative Institutions

University of Melbourne

Peter MacCallum Cancer Centre

Cancer Institute (WIA)

The Royal Melbourne Hospital

Walter and Eliza Hall Institute of Medical Research

Albert Einstein College of Medicine

Cancer Institute of New South Wales

National Cancer Institute

University of Sydney

Monash University

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Analysis of colonies developing in vitro from mouse bone marrow cells stimulated by kidney feeder layers or leukemic serum

Journal of Cellular Physiology (1967)

D Metcalf T. R. Bradley William A. Robinson

Abstract An analysis has been made of cell colonies developing in agar cultures from mouse bone marrow cells following stimulation either by neonatal kidney cell feeder layers or AKR lymphoid leukemia serum. Colonies arose by cell proliferation and were mixtures of granulocytic and mononuclear cells. Colonies stimulated by kidney feeder layers reached a mean size of 2000 cells by day 10 of incubation and remained predominantly granulocytic in nature. When bovine serum was substituted for fetal calf serum, cell colonies grew to a smaller size and lost their granulocytic nature, finally becoming almost pure populations of mononuclear cells. Colonies stimulated by AKR leukemic serum reached a mean size of 350 cells by day 10 of incubation. Although these colonies initially were granulocytic in nature, they finally became almost pure populations of mononuclear cells. The colony mononuclear cells actively phagocytosed carbon, and contained metachromatic granules probably derived from ingestion of agar. The mononuclear cells in these colonies may not have been members of the original colony, but may have been incorporated in the colony as it expanded in size, subsequently proliferating in the favourable environment of the colony.

10.1002/jcp.1040690112

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Citations (139)

Cytotoxicity of an 125I-labeled DNA-binding compound that induces double-stranded DNA breaks.

PubMed (1979)

Roger F. Martin T. R. Bradley G. S. Hodgson

[125I]Iodorivanol (6,9-diamino-2-ethoxy-5-[125]iodoacridine) has been prepared by direct iodination of rivanol (6.9-diamino-2-ethoxyacridine). In vitro binding of [125I]iodorivanol to PM2 DNA resulted in induction of double-stranded DNA breaks following decay of the 125I atom, presumably in the same way as decay of 125I atoms in 125I-labeled DNA causes double-stranded DNA breaks. Treatment of mouse L-cell cultures with [125I]iodorivanol resulted in a cell kill, the extent of which was dependent on the 125I specific activity and the duration of exposure. A clonogenic assay was used to quantitate cell kill. It was concluded that at least some of the [125I]iodorivanol in the culture medium was taken up by the cells, transported to the nucleus, and bound to DNA and that subsequent decay of the 125I atoms induced double-stranded DNA breaks in the genome, with consequent loss of viability. 125I-labeled DNA-binding compounds are suggested as a novel class of cytotoxic agents.

Clonogenic assay

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Progenitor cells in murine bone marrow stimulated by growth factors produced by the AF1-19T rat cell line.

PubMed (1990)

A. B. Kriegler T. R. Bradley I Bertoncello John A. Hamilton Prue H. Hart

Interleukin 3

Colony-stimulating factor

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Isolation and analysis of primitive hemopoietic progenitor cells on the basis of differential expression of Qa-m7 antigen.

The Journal of Immunology (1986)

Ivan Bertoncello SH Bartelmez T. R. Bradley E. Richard Stanley Robert A. Harris

In the presence of the hemopoietic growth factor CSF-1, the later committed cells of the macrophage lineage can be detected by their ability to form small colonies in clonal agar culture (CFCCSF-1). Synergistic factors have been described that in combination with CSF-1 stimulate developmentally early hemopoietic progenitor cells of high proliferative potential (HPP-CFC). By using a monoclonal antibody to the Qa-m7 antigenic determinant, we investigated and compared the expression of Qa-m7 on CFCCSF-1 and on HPP-CFC of two types that grow in response to either 1) CSF-1 plus synergistic factor from human placenta-conditioned medium (HPP-CFCHplac+CSF-1) or 2) CSF-1 plus synergistic factor from conditioned medium of the WEHI-3 myelomonocytic cell line (HPP-CFCW+CSF-1). We have shown that HPP-CFC of both types express relatively more Qa-m7 antigen than CFCCSF-1 and can be separated and enriched on this basis by discontinuous buoyant density centrifugation and fluorescence-activated cell sorting of normal bone marrow. Significant enrichments of HPP-CFCHPlac+CSF-1 (43.5-fold) and HPP-CFCW+CSF-1 (28.8-fold) have been achieved with cloning efficiencies of HPP-CFC in the most enriched fractions reaching 4 to 5%. These results clearly illustrate the fact that there are populations of progenitor cells from normal, unperturbed bone marrow that strictly require a combination of two hemopoietic growth factors (CSF-1 plus synergistic factor) in order to be detected.

Cell Sorting

Differential centrifugation

Colony-stimulating factor

10.4049/jimmunol.136.9.3219

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Bone Marrow Colony Stimulating Activity in Human Sera. RESULTS OF TWO INDEPENDENT SURVEYS IN BUFFALO AND MELBOURNE*

British Journal of Haematology (1968)

Roger S. Foster D Metcalf William A. Robinson T. R. Bradley

In two independent surveys, 1188 sera from 947 normal persons or patients with various diseases were tested for their capacity to stimulate mouse bone marrow cells to proliferate in vitro and form cell colonies. Ninety‐eight per cent of sera from normal persons had no colony stimulating activity. Colony stimulating activity was detected in sera from 15 to 70 per cent of patients with: (1) leukaemia and allied diseases, particularly in the advanced stages and active phases of the diseases, (2) proliferative diseases of leukopoiesis, and (3) non‐bacterial infections. Regardless of the source of the active serum, all colonies stimulated initially contained granulocytic cells and later contained phagocytic mononuclear cells.

Colony-stimulating factor

10.1111/j.1365-2141.1968.tb01524.x

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Citations (125)

Antigens expressed on human bone marrow cells, granulocyte-macrophage progenitors, myeloid and T cell lymphoblastic leukaemias

Pathology (1982)

G.R. Pilkington E. Januszewicz T. R. Bradley Ian A. Cooper David G. Jose

10.1016/s0031-3025(16)38326-x

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The Effect of Erythrocytes on Mouse Bone Marrow

T. R. Bradley P. A. Telfer Phyllis Fry

The medium used was Eagle’s Minimal Essential Medium supplemented with pyruvate ( 1 mM ), serine (0.2 mM), soy broth ( 10% ), fetal calf serum ( 10% ) and horse serum ( 5 % ) 4 being made up at x 2 concentration and mixed with equal volumes of 1 % agar for underlays ( 5 ml per 60 mm Petri dish) or 0.6% agar for overlays (2 ml per dish) prior to plating. For stimulation of colony formation a large batch of partially purified extract of mouse embryos was prepared by a slight variation of a method previously described.4 Frozen whole mouse embryos (BALB/c, 16th to 20th clay of pregnancy) were homogenized in a \\Taring Blendor with 3 volumes of cold distilled water. After centrifugation at 15,000 g for 30 mm, the supernatant was brought to 50% saturation with ammonium sulfate and the precipitate removed by centrifugation (15,000 g for 30 mm). The supernatant was then

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Distilled water

Ammonium sulfate

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Human cell cultures for screening anti-cancer drugs

Trends in Pharmacological Sciences (1987)

Ivan Bertoncello T. R. Bradley

10.1016/0165-6147(87)90193-3

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Citations (5)