scholarly journals A HISTOCHEMICAL EVALUATION OF THE PYROANTIMONATE-OSMIUM REACTION

1971 ◽  
Vol 19 (12) ◽  
pp. 727-737 ◽  
Author(s):  
MICHAEL A. CLARK ◽  
G. ADOLPH ACKERMAN
Keyword(s):  
Bone Marrow Cells ◽  
Organometallic Complexes ◽  
Control Measures ◽  
Amino Groups ◽  
Metallic Cation ◽  
Normal Human ◽  
Hydroxy Groups ◽  
Nucleolar Staining ◽  
Acid Mucosubstances ◽  
Normal Human Bone Marrow

The chemical nature of the pyroantimonate-osmium (PAO) reaction in normal human bone marrow cells has been evaluated by the application of a number of digestion and blocking procedures as well as the electron microprobe. Five foci of reactivity are localized in cells fixed directly in the PAO reagent, viz., nucleoli, heterochromatin, cytoplasmic granules, particulate glycogen and the outer plasmalemmal surface. Heterochromatin and nucleolar staining are attributable to calcium bound to nucleic acids as well as to reactive amino groups on histones. Granule staining is dissociable into acid-dialyzable, ribonuclease (RNase)-stable and RNase-labile components but sulfate groups of acid mucosubstances are probably not involved. Glycogen staining by the PAO reagent has been verified and shown to result from the formation of organometallic complexes between the pyroantimonate ion and the C2-C3-free hydroxy groups of the glucose residues. PAO reactivity has been definitely localized along the outer plasmalemmal surface, but this component of PAO staining is resistant to all control measures employed. These studies have illustrated the complexity and polyvalency of the PAO reaction and have shown that PAO staining is not exclusively associated with metallic cation localization.

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

Blood ◽  
1996 ◽  
Vol 87 (3) ◽  
pp. 977-982 ◽  
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.

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

1979 ◽  
Vol 50 (2) ◽  
pp. 213-216 ◽  
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

Nitric oxide induces apoptosis in megakaryocytic cell lines

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3451-3459 ◽  
Author(s):  
Elisabeth Battinelli ◽  
Joseph Loscalzo
Keyword(s):  
Nitric Oxide ◽  
Cell Lines ◽  
Bone Marrow Cells ◽  
No Synthase ◽  
Cell Systems ◽  
Normal Human ◽  
Induced Apoptosis ◽  
Normal Human Bone Marrow ◽  
Megakaryocytic Cell ◽  
Inducible Nitric Oxide

Cytokines that stimulate inducible nitric oxide (NO) synthase can suppress the growth and differentiation of normal human bone marrow cells, including megakaryocytes. Since NO promotes apoptosis in other cell systems, we chose to study the determinants of apoptosis in megakaryocytic cells. We show that both exogenous and endogenous sources of NO can induce apoptosis in megakaryocytoid cell lines. The megakaryocyte growth factor thrombopoietin suppresses NO-induced apoptosis, whereas treatment with peroxynitrite, a cytotoxic product formed when NO reacts with superoxide, promotes apoptosis. Superoxide inhibitors suppress NO-induced apoptosis, and pretreatment with megakaryocyte growth and maturation factors attenuates NO-induced apoptosis. These data show that NO modulates megakaryocyte apoptosis and suggest that this process may occur in the cytokine-rich marrow milieu to regulate megakaryocyte turnover.

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

Blood ◽  
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.

scholarly journals Expression of the YB5.B8 antigen (c-kit proto-oncogene product) in normal human bone marrow

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 30-37 ◽  
Author(s):  
LK Ashman ◽  
AC Cambareri ◽  
LB To ◽  
RJ Levinsky ◽  
CA Juttner
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Lymphoid Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Stem ◽  
Normal Peripheral Blood ◽  
Normal Human ◽  
Normal Human Bone Marrow

Abstract The c-kit proto-oncogene product is a member of the family of growth factor receptors with intrinsic tyrosine kinase activity. In the mouse c-kit maps to the W locus, which is known to be of central importance in hematopoiesis. Monoclonal antibody (MoAb) YB5.B8, which was raised against peripheral blood blast cells from a patient with acute myeloid leukemia (AML), was recently shown to bind to the extracellular domain of the c-kit product. This antibody does not bind detectably to normal peripheral blood cells and identifies a sub-group of AML patients with poor prognosis. We have used MoAb YB5.B8 to study the expression of c- kit by normal human bone marrow cells by immunofluorescence and flow cytometry, and to isolate multipotential and erythroid colony-forming cells. In a series of 11 normal adult bone marrow specimens, MoAb YB5.B8 bound to 4.0% +/- 1.8% of the cells in the low-density fraction. Dual-labeling experiments were performed with YB5.B8, and CD33, CD34, and CD10 MoAbs. Three populations of cells binding YB5.B8 could be identified based on their pattern of coexpression of the other markers; ie, YB5.B8+/CD34+/CD33-, YB5.B8+/CD34+/CD33+ and YB5.B8+/CD34+/CD33+. These populations had distinctive two-dimensional light scatter characteristics and are likely to correspond to precursor colony- forming cells, colony-forming cells, and maturing mast cells, respectively. No cells binding both YB5.B8 and an MoAb to the early lymphoid marker CD10 were found, implying that most early lymphoid cells do not express c-kit. MoAbs to the c-kit protein should prove valuable in multimarker studies of human hematopoietic stem and progenitor cells. Definition of a reference range of c-kit expression in normal human bone marrow will provide a sound basis for further studies of this marker in diagnosis and prognosis in AML.

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