The Influence of Recombinant Human Granulocyte-Monocyte-Colony-Stimulating Factor on Stroma Formation by Normal Human Bone Marrow Cells In Vitro

Zidovudine, the antiviral drug used in the treatment of acquired immunodeficiency syndrome (AIDS), causes toxicity to the haematopoietic system. Although use of the haematopoietic growth factors, GM-CSF and erythropoietin have been investigated in clinical trials to modulate antiviral toxicity, there is scant data which supports their ability to ameliorate zidovudine induced toxicity on haematopoietic progenitor cells when combined in vitro. We describe here the results of studies designed to evaluate the capacity of additional haematopoietic factors such as granulocyte-colony stimulating factor (G-CSF) and macrophage-colony stimulating factor (M-CSF) to modulate zidovudine-induced toxicity on G-CSF and M-CSF dependent-colony formation in the presence or absence of zidovudine in vitro. These factors were also studied combined with erythropoietin in culture for the early erythroid progenitor BFU-E using adherent, T-cell, depleted normal human bone marrow cells in the presence or absence of zidovudine. In the presence of zidovudine at the concentration producing 50% inhibition of G- and M-CSF dependent colony formation, (5 × 10−5M), dose-escalation of either G-CSF or M-CSF failed to ameliorate zidovudine toxicity. However, in the presence of zidovudine at the concentration that produces 50% inhibition of BFU-E (5 × 10−9M), and optimal erythropoietin (1 unit ml−1), G-CSF ameliorated zidovudine inhibition of BFU-E, which was not observed with M-CSF. In the presence of erythropoietin, G-CSF increased significantly normal BFU-E. These studies indicate that G-CSF may be useful in ameliorating zidovudine-induced anaemia and suggest G-CSF may act as a synergistic factor to enhance erythropoietin to support the growth of erythroid progenitors in conditions where erythropoitin is ineffective.

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Fractionation of subsets of BFU-E from normal human bone marrow: responsiveness to erythropoietin, human placental-conditioned medium, or granulocyte-macrophage colony-stimulating factor

Blood ◽

10.1182/blood.v71.3.758.758 ◽

1988 ◽

Vol 71 (3) ◽

pp. 758-765 ◽

Cited By ~ 20

Author(s):

G Kannourakis ◽

GR Johnson

Keyword(s):

Bone Marrow ◽

Conditioned Medium ◽

Human Bone ◽

Human Bone Marrow ◽

Colony Stimulating Factor ◽

Normal Human ◽

Macrophage Colony Stimulating Factor ◽

Macrophage Colony ◽

Stimulating Factor ◽

Normal Human Bone Marrow

Abstract Normal human bone marrow mononuclear cells were fractionated by differential adherence, immunomagnetic separation, and fluorescence- activated cell sorting (FACS). The resultant fractionated cells were cultured in semisolid medium to monitor the presence of BFU-E, Mix-CFC, and nonerythroid CFC. Two populations of cells were recovered on the basis of binding by the monoclonal antibody (MoAb) RFB-1. One of these populations contained BFU-E that were stimulated only by erythropoietin (Epo), whereas the second population contained BFU-E responsive to Epo, Epo and recombinant human granulocyte-macrophage colony-stimulating factor (rHGM-CSF), or Epo and human placental-conditioned medium (HPCM). Prior enrichment of clonogenic cells by removal of adherent and Leu-M3+ve, Leu-4+ve, Leu-7+ve, B1+ve, WEMG1+ve, and Glycophorin A+ve cells, followed by FACS fractionation on the basis of RFB-1 binding, consistently resulted in recoveries of BFU-E, Mix-CFC, and nonerythroid CFC of greater than 100% (up to 800%). These procedures also resulted in enrichment of up to 200-fold and frequencies of 1:6 for BFU-E, 1:5 for CFC, and 1:130 for Mix-CFC.

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Ultrastructural localization of peroxidase activity in normal human bone marrow cells

Cell and Tissue Research ◽

10.1007/bf00330708 ◽

1971 ◽

Vol 117 (4) ◽

pp. 463-475 ◽

Cited By ~ 34

Author(s):

G. Adolph Ackerman ◽

Michael A. Clark

Keyword(s):

Bone Marrow ◽

Peroxidase Activity ◽

Bone Marrow Cells ◽

Ultrastructural Localization ◽

Human Bone ◽

Human Bone Marrow ◽

Normal Human ◽

Normal Human Bone Marrow ◽

Marrow Cells

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Differential effects of nitric oxide on erythroid and myeloid colony growth from CD34+ human bone marrow cells

Blood ◽

10.1182/blood.v87.3.977.bloodjournal873977 ◽

1996 ◽

Vol 87 (3) ◽

pp. 977-982 ◽

Cited By ~ 56

Author(s):

PJ Shami ◽

JB Weinberg

Keyword(s):

Bone Marrow ◽

Mononuclear Cells ◽

Bone Marrow Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Monocytic Differentiation ◽

Colony Forming Unit ◽

Normal Human ◽

Normal Human Bone Marrow ◽

Marrow Cells

Nitric oxide (NO) is a reactive molecule with numerous physiologic and pathophysiologic roles affecting the nervous, cardiovascular, and immune systems. In previous work, we have demonstrated that NO inhibits the growth and induces the monocytic differentiation of cells of the HL- 60 cell line. We have also demonstrated that NO inhibits the growth of acute nonlymphocytic leukemia cells freshly isolated from untreated patients and increases monocytic differentiation antigens in some. In the present work, we studied the effect of NO on the growth and differentiation of normal human bone marrow cells in vitro. Mononuclear cells isolated from human bone marrow were cultured in semisolid media and treated with the NO-donating agents sodium nitroprusside (SNP) or S- nitroso-acetyl penicillamine (SNAP) (0.25 to 1 mmol/L). Both agents decreased colony-forming unit-erythroid (CFU-E) and colony-forming unit- granulocyte macrophage (CFU-GM) formation by 34% to 100%. When CD34+ cells were examined, we noted that these cells responded to SNP and SNAP differently than did the mononuclear cells. At a concentration range of 0.25 to 1 mmol/L, SNP inhibited the growth of CFU-E by 30% to 75%. However, at the same concentration range, SNP increased the number of CFU-GM by up to 94%. At concentrations of 0.25 to 1 mmol/L, SNAP inhibited the growth of CFU-E by 33% to 100%. At a concentration of 0.25 mmol/L, SNAP did not affect CFU-GM. At higher concentrations, SNAP inhibited the growth of CFU-GM. Although SNP increased intracellular levels of cGMP in bone marrow cells, increasing cGMP in cells by addition of 8-Br-cGMP (a membrane permeable cGMP analogue) did not reproduce the observed NO effects on bone marrow colonies. These results demonstrate that NO can influence the growth and differentiation of normal human bone marrow cells. NO (generated in the bone marrow microenvironment) may play an important role modulating the growth and differentiation of bone marrow cells in vivo.

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The rate of sister chromatid exchange in normal human bone marrow cells

Human Genetics ◽

10.1007/bf00390243 ◽

1979 ◽

Vol 50 (2) ◽

pp. 213-216 ◽

Cited By ~ 17

Author(s):

R. Becher ◽

C. G. Schmidt ◽

Gabriele Theis ◽

D. K. Hossfeld

Keyword(s):

Bone Marrow ◽

Sister Chromatid ◽

Sister Chromatid Exchange ◽

Bone Marrow Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Normal Human ◽

Normal Human Bone Marrow ◽

Marrow Cells

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Phenotypic variations dictate the intracellular compartmentalization of doxorubicin in normal human bone marrow cells

Cancer Chemotherapy and Pharmacology ◽

10.1007/bf00684889 ◽

1990 ◽

Vol 25 (4) ◽

pp. 295-298 ◽

Cited By ~ 3

Author(s):

Esther Agha� ◽

Zolt�n A. T�k�s

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Normal Human ◽

Phenotypic Variations ◽

Normal Human Bone Marrow ◽

Intracellular Compartmentalization ◽

Marrow Cells

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Proliferation of diffusion chamber colony-forming units (CFUD) in cultures of normal human bone marrow in diffusion chambers in mice

Blood ◽

10.1182/blood.v49.3.415.bloodjournal493415 ◽

1977 ◽

Vol 49 (3) ◽

pp. 415-424

Author(s):

N Jacobsen

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Diffusion Chambers ◽

Colony Forming Units ◽

Normal Human ◽

Normal Human Bone Marrow

Normal human bone marrow contains cells capable of forming colonies of hemopoietic cells in fibrin clots in diffusion chambers implanted intraperitoneally (i.p.) into irradiated mice. The present paper describes the proliferation of such colony-forming units (CFUD) in cultures in vivo. Cells harvested from diffusion chambers after 1–14 days of culture in 450-R irradiated mice contained CFUD, which formed neutrophilic, eosinophilic, or megakaryocytic colonies when tested by secondary culture in fibrin clot chambers. When bone marrow was precultured in irradiated mice at a concentration of 10(6) cells per chamber, an initial fall in the number of neutrophilic CFUD was observed. This decrease was followed by an increase to a maximum at day 2, and then a secondary decrease. The number of neutrophilic CFUD recovered after 2 days of preculture in irradiated mice varied between 60% and 250% of the number present before preculture. Preculture in nonirradiated mice resulted in a significantly lower recovery of neutrophilic CFUD. In vitro treatment of bone marrow cells with hydroxyurea (OHU) after 2 days of preculture in irradiated mice resulted in a 68% +/- 5% reduction in the number of neutrophilic CFUD. In contrast, OHU had no similar effect on precultures from nonirradiated mice. Both the recovery and sensitivity to OHU of eosinophilic CFUD were independent of host irradiation. Similarly, no effect of host irradiation on the recovery or the 3H-thymidine (3HTdR) labeling index of morphologically recognizable granulocytic cells was observed at day 2. The data suggest an effect of humoral host factor(s) on the proliferation of early precursor cells, which are or become committed to differentiate into the neutrophilic pathway in diffusion chambers.

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