scholarly journals Stimulation of proliferation of human myeloid leukemia cells in culture: applications for cytogenetic analysis

Blood ◽  
1986 ◽  
Vol 68 (3) ◽  
pp. 790-793 ◽  
Author(s):  
J Michaeli ◽  
I Lerer ◽  
EA Rachmilewitz ◽  
E Fibach
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Peripheral Blood ◽  
Cytogenetic Analysis ◽  
Bone Marrow Cells ◽  
Leukemic Cells ◽  
Pronounced Increase ◽  
Stimulation Of ◽  
Marrow Cells ◽  
Mitotic Cells

Abstract The use of chromosome banding techniques has provided a valuable diagnostic tool in various malignancies. The application of these methods, however, is often restricted by a low yield of mitotic cells and the patient's unwillingness to comply with repeated bone marrow aspiration. In an attempt to promote mitotic activity of leukemic cells from the bone marrow and peripheral blood, we employed a new method based on culturing the cells in the presence of a conditioned medium derived from a human bladder carcinoma cell line (5637). In addition to colony stimulating factor, this conditioned medium contains a factor that is capable of stimulating leukemic myeloblast proliferation. Bone marrow and peripheral blood mononuclear cells from 58 patients with a variety of myeloid leukemias were cultured for 24 to 120 hours in the presence or absence of conditioned medium. These bone marrow cells showed a pronounced increase in the mitotic index (5- to 50-fold) as compared to unstimulated cultures, and a greater than 100-fold increase as compared to fresh, uncultured bone marrow cells. Analyzable metaphases could be obtained even in marrow samples in which direct or 24-hour G-banding techniques had failed to reveal metaphases. The effect observed on peripheral blood cells was even more dramatic because prior to culture no mitotic cells were detected, whereas up to 2% mitotic cells were found in conditioned medium-stimulated peripheral leukemic cells. Karyotype analysis of 36 out of the 58 leukemic patients has shown that the chromosome changes discovered in conditioned medium stimulated cells were identical to those found in unstimulated cells. New chromosome aberrations, attributable to the stimulation of growth by conditioned medium, were not found. The quality of the metaphases analyzed following conditioned medium stimulation was considerably better than that of unstimulated samples. Frozen cells, when cultured with conditioned medium, were also suitable for cytogenetic analysis. Thus, the use of this conditioned medium permits adequate cytogenetic analysis even in cases where such analysis was previously impossible.

scholarly journals Stimulation of proliferation of human myeloid leukemia cells in culture: applications for cytogenetic analysis

Blood ◽  
1986 ◽  
Vol 68 (3) ◽  
pp. 790-793
Author(s):  
J Michaeli ◽  
I Lerer ◽  
EA Rachmilewitz ◽  
E Fibach
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Peripheral Blood ◽  
Cytogenetic Analysis ◽  
Bone Marrow Cells ◽  
Leukemic Cells ◽  
Pronounced Increase ◽  
Stimulation Of ◽  
Marrow Cells ◽  
Mitotic Cells

The use of chromosome banding techniques has provided a valuable diagnostic tool in various malignancies. The application of these methods, however, is often restricted by a low yield of mitotic cells and the patient's unwillingness to comply with repeated bone marrow aspiration. In an attempt to promote mitotic activity of leukemic cells from the bone marrow and peripheral blood, we employed a new method based on culturing the cells in the presence of a conditioned medium derived from a human bladder carcinoma cell line (5637). In addition to colony stimulating factor, this conditioned medium contains a factor that is capable of stimulating leukemic myeloblast proliferation. Bone marrow and peripheral blood mononuclear cells from 58 patients with a variety of myeloid leukemias were cultured for 24 to 120 hours in the presence or absence of conditioned medium. These bone marrow cells showed a pronounced increase in the mitotic index (5- to 50-fold) as compared to unstimulated cultures, and a greater than 100-fold increase as compared to fresh, uncultured bone marrow cells. Analyzable metaphases could be obtained even in marrow samples in which direct or 24-hour G-banding techniques had failed to reveal metaphases. The effect observed on peripheral blood cells was even more dramatic because prior to culture no mitotic cells were detected, whereas up to 2% mitotic cells were found in conditioned medium-stimulated peripheral leukemic cells. Karyotype analysis of 36 out of the 58 leukemic patients has shown that the chromosome changes discovered in conditioned medium stimulated cells were identical to those found in unstimulated cells. New chromosome aberrations, attributable to the stimulation of growth by conditioned medium, were not found. The quality of the metaphases analyzed following conditioned medium stimulation was considerably better than that of unstimulated samples. Frozen cells, when cultured with conditioned medium, were also suitable for cytogenetic analysis. Thus, the use of this conditioned medium permits adequate cytogenetic analysis even in cases where such analysis was previously impossible.

scholarly journals Normal Granulocyte Colony-forming Cells in the Bone Marrow of Yemenite Jews With Genetic Neutropenia

Blood ◽  
1973 ◽  
Vol 41 (6) ◽  
pp. 745-751 ◽  
Author(s):  
Uri Mintz ◽  
Leo Sachs
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Human Embryo ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Release Mechanism ◽  
Neutropenic Patients ◽  
Yemenite Jews ◽  
Marrow Cells

Abstract The frequency of normal granulocyte colony-forming cells in the bone marrow of Yemenite Jews with genetic, absolute neutropenia and no special tendency to infection has been studied with conditioned medium from human embryo fibroblasts. Cells from these neutropenic patients gave an average of about twice the number of granulocyte colonies as bone marrow cells from nonneutropenic patients. No morphologic abnormalities were observed in the granulocytes in bone marrow smears or in colonies formed in vitro. The results indicate that the neutropenia in these patients was not due to a deficiency of granulocyte colony-forming cells. It is suggested that the neutropenia is due to a defect in the release mechanism of mature granulocytes from the bone marrow to the peripheral blood.

Limits of stimulation of proliferation and differentiation of bone marrow cells of mice treated with swainsonine

2003 ◽  
Vol 3 (10-11) ◽  
pp. 1537-1547 ◽  
Author(s):  
Oladipo A. Oredipe ◽  
Paulette M. Furbert-Harris ◽  
Ibrahim Laniyan ◽  
Walter M. Griffin ◽  
Rajagopalan Sridhar
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Proliferation And Differentiation ◽  
Stimulation Of ◽  
Marrow Cells

scholarly journals Adult Mice With Targeted Mutation of the Interleukin-11 Receptor (IL11Ra) Display Normal Hematopoiesis

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2148-2159 ◽  
Author(s):  
Harshal H. Nandurkar ◽  
Lorraine Robb ◽  
David Tarlinton ◽  
Louise Barnett ◽  
Frank Köntgen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Bone Marrow Cells ◽  
White Blood Cells ◽  
Null Mutation ◽  
Wild Type ◽  
Normal Numbers ◽  
Interleukin 11 ◽  
Marrow Cells

Abstract Interleukin-11 (IL-11) is a pleiotropic growth factor with a prominent effect on megakaryopoiesis and thrombopoiesis. The receptor for IL-11 is a heterodimer of the signal transduction unit gp130 and a specific receptor component, the α-chain (IL-11Rα). Two genes potentially encode the IL-11Rα: the IL11Ra and IL11Ra2 genes. The IL11Ra gene is widely expressed in hematopoietic and other organs, whereas the IL11Ra2 gene is restricted to only some strains of mice and its expression is confined to testis, lymph node, and thymus. To investigate the essential actions mediated by the IL-11Rα, we have generated mice with a null mutation of IL11Ra (IL11Ra−/−) by gene targeting. Analysis of IL11Ra expression by Northern blot and reverse transcriptase-polymerase chain reaction, as well as the absence of response of IL11Ra−/− bone marrow cells to IL-11 in hematopoietic assays, further confirmed the null mutation. Compensatory expression of the IL11Ra2 in bone marrow cells was not detected. IL11Ra−/− mice were healthy with normal numbers of peripheral blood white blood cells, hematocrit, and platelets. Bone marrow and spleen contained normal numbers of cells of all hematopoietic lineages, including megakaryocytes. Clonal cultures did not identify any perturbation of granulocyte-macrophage (GM), erythroid, or megakaryocyte progenitors. The number of day-12 colony-forming unit-spleen progenitors were similar in wild-type and IL11Ra−/− mice. The kinetics of recovery of peripheral blood white blood cells, platelets, and bone marrow GM progenitors after treatment with 5-flurouracil were the same in IL11Ra−/− and wild-type mice. Acute hemolytic stress was induced by phenylhydrazine and resulted in a 50% decrease in hematocrit. The recovery of hematocrit was comparable in IL11Ra−/− and wild-type mice. These observations indicate that IL-11 receptor signalling is dispensable for adult hematopoiesis.

scholarly journals Analysis of interferon-inducible genes in cells of chronic myeloid leukemia patients responsive or resistant to an interferon-alpha treatment

Blood ◽  
1990 ◽  
Vol 76 (11) ◽  
pp. 2337-2342
Author(s):  
IM Clauss ◽  
B Vandenplas ◽  
MG Wathelet ◽  
C Dorval ◽  
A Delforge ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Myeloid Leukemia ◽  
Interferon Alpha ◽  
Peripheral Blood ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Healthy Controls ◽  
Ifn Alpha ◽  
Inducible Genes ◽  
Marrow Cells

Recombinant human interferon-alpha (IFN-alpha) can induce a hematologic remission in patients with chronic myeloid leukemia. However, some patients are resistant and others develop late resistance to the IFN- alpha treatment. To understand the molecular mechanism of this resistance, we have analyzed the expression of 10 IFN-inducible genes in the cells of three resistant patients, two responsive patients, and six healthy controls. Northern blot hybridizations showed that all the genes were induced in in vitro IFN-alpha treated peripheral blood cells of the patients and healthy controls. These genes were also inducible in peripheral blood and bone marrow cells of two out of two resistant patients administered an injection of IFN-alpha. We conclude that the resistance to the IFN-alpha treatment of the chronic myeloid leukemia patients we studied is not due to (1) the absence of induction of any of the 10 IFN-inducible genes we studied, including the low-molecular- weight 2′-5′oligoadenylate synthetase; (2) the presence of an antagonist of IFN-alpha in the peripheral blood or bone marrow cells; and (3) the presence of neutralizing anti-IFN-alpha antibodies.

scholarly journals In vitro stimulation of primate hemoglobin synthesis by L-thyroxine

Blood ◽  
1977 ◽  
Vol 49 (3) ◽  
pp. 407-413 ◽  
Author(s):  
JE Fuhr ◽  
N Gengozian ◽  
M Overton
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Leucine Incorporation ◽  
Hemoglobin Synthesis ◽  
Globin Chains ◽  
Stimulation Of ◽  
Marrow Cells

Abstract Bone marrow cells from adult and abortus primates (marmosets) were incubated in vitro to determine their responsiveness to L-thyroxine. 3H- leucine incorporation into purified globin chains was the parameter assayed to determine responsiveness. Bone marrow from spontaneously aborted animals consistently was stimulated by the presence of physiologic levels of L-thyroxine. Bone marrow cells from adult animals were unaffected by the hormone.

scholarly journals 5-Azacytidine increases gamma-globin synthesis and reduces the proportion of dense cells in patients with sickle cell anemia

Blood ◽  
1983 ◽  
Vol 62 (2) ◽  
pp. 370-380 ◽  
Author(s):  
TJ Ley ◽  
J DeSimone ◽  
CT Noguchi ◽  
PH Turner ◽  
AN Schechter ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Beta Thalassemia ◽  
Globin Genes ◽  
Gamma Globin ◽  
Globin Synthesis ◽  
Marrow Cells

Abstract We previously demonstrated that 5-azacytidine can selectively increase gamma-globin synthesis in a patient with beta +-thalassemia, prompting us to treat two patients with sickle cell anemia and two additional patients with beta + thalassemia. 5-Azacytidine (2 mg/kg/day) was continuously infused for 7 days with no apparent clinical toxicity. The gamma/beta-globin biosynthetic ratio increased fourfold to sixfold in the bone marrow cells of each patient after treatment and remained elevated for 7–14 additional days. Hypomethylation of DNA near the gamma-globin genes in bone marrow cells was demonstrated 2 days after beginning the 5-azacytidine infusion. The peripheral blood fetal hemoglobin (HbF) level increased from 6.0% to 13.7% in one patient with sickle cell anemia and from 1.6% to 8.9% in the second. Stractan gradient analyses of peripheral blood from patients with sickle cell anemia revealed a marked decrease in the percentage of dense cells (cells that contain increased amounts of HbS polymer when deoxygenated) following treatment. These observations provide an impetus to investigate the effects of repeated courses of 5-azacytidine in a small group of severely ill patients to determine whether this drug may have a role in the treatment of patients with sickle cell anemia and beta- thalassemia.

scholarly journals Detection by enzymatic amplification of bcr-abl mRNA in peripheral blood and bone marrow cells of patients with chronic myelogenous leukemia

Blood ◽  
1989 ◽  
Vol 73 (6) ◽  
pp. 1735-1741 ◽  
Author(s):  
W Lange ◽  
DS Snyder ◽  
R Castro ◽  
JJ Rossi ◽  
KG Blume
Keyword(s):  
Bone Marrow ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Philadelphia Chromosome ◽  
Chromosome 9 ◽  
Myelogenous Leukemia ◽  
Enzymatic Amplification ◽  
Polymerase Chain ◽  
Marrow Cells

Abstract The Philadelphia chromosome of chronic myelogenous leukemia (CML) patients is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. A new bcr- abl mRNA is expressed in these cases. We have developed a modified polymerase chain reaction (PCR) for the detection of this mRNA. The method is extremely sensitive, reliable, and relatively fast. The analysis of peripheral blood or bone marrow cells from CML patients treated with chemotherapy shows that the two possible mRNAs are expressed in various combinations. Our results show that even after myeloablative therapy for bone marrow transplantation bcr-abl mRNAs are still expressed. Further studies, however, are necessary to determine the clinical relevance of a small number of persisting cells expressing the bcr-abl mRNA.

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