scholarly journals Effect of Erythropoietin on RNA Synthesis by Normal and Leukemic Bone Marrow and Spleen Cell Suspensions In Vitro

Blood ◽  
1974 ◽  
Vol 44 (4) ◽  
pp. 535-542 ◽  
Author(s):  
Evelyn E. Handler ◽  
Naomi Mendelsohn ◽  
Eugene S. Handler
Keyword(s):  
Bone Marrow ◽  
Rna Synthesis ◽  
Bone Marrow Cells ◽  
Marrow Cell ◽  
Cell Suspensions ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Spleen Cells ◽  
Normal Bone Marrow Cell

Abstract Erythropoietin (EPO) induced a 42% increase in 3H-uridine incorporation into RNA after a 5-hr culture of normal bone marrow cell suspensions. Bone marrow cells obtained from rats 3-5 days after the initiation of a myelogenous leukemia exhibited a decreased responsivity to EPO. At this time incorporation of the isotope into RNA in the presence of EPO was approximately 50% of controls. Rats rendered leukemic 8-10 days prior to culture showed no bone marrow response to EPO even in those instances where leukemic cells comprised a relatively small percentage of the marrow compartment. EPO had little or no effect on RNA synthesis by spleen cells obtained from normal and leukemic rats. This was noted even in those leukemic spleens in which erythropoiesis was observed. The data suggest that the anemia associated with myelogenous leukemia may, in part, be due to a loss of EPO-responsive cells and/or a loss of sensitivity of these elements to normal humoral control.

scholarly journals Colony formation in vitro by leukemic cells in acute lymphoblastic leukemia (ALL)

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 712-718 ◽  
Author(s):  
SD Smith ◽  
EM Uyeki ◽  
JT Lowman
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Bone Marrow Cells ◽  
Lymphoblastic Leukemia ◽  
Lymphoid Cells ◽  
Assay System ◽  
Leukemic Cells ◽  
Specific Cell ◽  
Specific Cell Surface

Abstract An assay system in vitro for the growth of malignant lymphoblastic colony-forming cells (CFC) was established. Growth of malignant myeloblastic CFC has been previously reported, but this is the first report of growth of malignant lymphoblastic CFC. Established assay systems in vitro have been very helpful in elucidating the control of growth and differentiation of both normal and malignant bone marrow cells. Lymphoblastic CFC were grown from the bone marrow aspirates of 20 children with acute lymphoblastic leukemia. Growth of these colonies was established on an agar assay system and maintained in the relative hypoxia (7% oxygen) of a Stulberg chamber. The criteria for malignancy of these colonies was based upon cellular cytochemical staining characteristics, the presence of specific cell surface markers, and the ability of these lymphoid cells to grow without the addition of a lymphoid mitogen. With this technique, specific nutritional requirements and drug sensitivities can be established in vitro, and these data may permit tailoring of individual antileukemic therapy.

scholarly journals Cellular maturation in human preleukemia

Blood ◽  
1978 ◽  
Vol 52 (2) ◽  
pp. 355-361
Author(s):  
HP Koeffler ◽  
DW Golde
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
Liquid Culture ◽  
Cultured Cells ◽  
Bone Marrow Cells ◽  
Myelogenous Leukemia ◽  
Bone Marrow Cultures ◽  
Acute Myelogenous ◽  
Marrow Cultures

Bone marrow cells from three preleukemic patients with prominent marrow karyotypic abnormalities were studied in liquid culture to determine if the neoplastic clones were capable of maturation. Parallel cytogenetic and cytologic studies were performed in sequentially harvested bone marrow cultures. Maturation, albeit delayed, occurred in cultures from all three patients. By 14 days of culture in vitro, morphologic, cytochemical, and functional evidence of maturation was observed in about 70% of the cells. By day 21, 85% of the cells were mature by these criteria. All but 2 of 249 metaphases from the cultured cells contained the cytogenetic abnormality of the neoplastic clone. We conclude that some preleukemic cells identified by a chromosomal abnormality can mature in vitro. Preleukemia may be viewed as a syndrome of “early leukemia” in which the neoplastic clone is established and manifested functionally as ineffective hematopoiesis. Hematopoietic cell differentiation becomes progressively abnormal with termination in the nearly complete maturational block characteristic of acute myelogenous leukemia.

scholarly journals In vitro restoration of polyclonal hematopoiesis in a chronic myelogenous leukemia after in vitro treatment with 4- hydroperoxycyclophosphamide

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 753-757 ◽  
Author(s):  
G Degliantoni ◽  
L Mangoni ◽  
V Rizzoli
Keyword(s):  
Bone Marrow ◽  
Chronic Myelogenous Leukemia ◽  
Early Phase ◽  
Bone Marrow Cells ◽  
Type A ◽  
G6pd Activity ◽  
Myelogenous Leukemia ◽  
Stem Cell Population ◽  
Marrow Cells

Bone marrow cells of a 45-year-old female with Philadelphia chromosome (Ph1)-positive, early-phase chronic myelogenous leukemia (CML), who was heterozygous for the glucose-6-phosphate dehydrogenase (G6PD) locus, were pretreated in vitro with 4-hydroperoxycyclophosphamide (4-HC) and tested for G6PD activity in several colony formation assays and for karyotypic abnormalities. All cells within the mixed (CFU-GEMM), the erythroid burst (BFU-E), and the granulocyte-macrophage (CFU-GM) colonies expressed type A and type B G6PD activity and a normal karyotype, whereas untreated cells expressed type A G6PD and the Ph1 chromosome. This reversal of G6PD activity type and the disappearance of the Ph1 chromosome in colonies grown from 4-HC-treated cells indicate that this cytotoxic agent spares a residual normal stem cell population in bone marrow cells of early-phase CML patients. This finding, in turn, suggests a therapeutic approach in CML based on in vitro chemotherapy of autologous bone marrow grafts.

scholarly journals Accelerated myeloblast destruction and abnormal lysosome disruption in cultured bone marrow: association with indolent acute myelogenous leukemia

Blood ◽  
1976 ◽  
Vol 48 (1) ◽  
pp. 23-32 ◽  
Author(s):  
RF Branda ◽  
HS Jacob ◽  
SD Douglas ◽  
CF Moldow ◽  
RR Puumala
Keyword(s):  
Bone Marrow ◽  
Acute Myelogenous Leukemia ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Electron Microscopic ◽  
Granule Formation ◽  
Microscopic Studies ◽  
Acute Myelogenous ◽  
Primary Granules

Abstract Despite no chemotherapy and a marrow morphologically typical of frank relapse, an acute myelogenous leukemia (AML) patient survived for nearly 1 yr. During this time she remained asymptomatic and maintained nearly normal levels of platelets and hemoglobin. Cytochemical and electron microscopic studies of her bone marrow in liquid culture revealed on several occasions a unique maturational sequence in that leukemic cells differentiated to form morphologically abnormal primary granules which appeared to rupture and cause cytolysis of these cells. In these cultures, blasts rapidly disappeared and were replaced by more mature granulocytes, in contrast to observations in cultures derived from five other patients with AML in relapse which showed persistently elevated blast counts with no evidence of maturation in vitro. These findings support the concept that in AML cell maturation is regularly impaired and in some cases also aberrant. In addition, the abnormal granule formation with autolysis of the leukemic cells observed in one patient may explain both the early cell death in vitro and this patient's relatively indolent clinical course. Similar in vitro studies may help predict atypical clinical courses in patients with AML and facilitate design of appropriate chemotherapy.

scholarly journals Interleukin-1 (22-K factor) induces release of granulocyte-macrophage colony-stimulating activity from human mononuclear phagocytes

Blood ◽  
1986 ◽  
Vol 68 (6) ◽  
pp. 1316-1321 ◽  
Author(s):  
WE Fibbe ◽  
J van Damme ◽  
A Billiau ◽  
PJ Voogt ◽  
N Duinkerken ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cell ◽  
Marrow Cell ◽  
Interleukin 1 ◽  
Colony Growth ◽  
Cell Suspensions ◽  
Mononuclear Phagocytes ◽  
Adherent Cells ◽  
Macrophage Colony

Abstract An electrophoretically pure preparation of natural human interleukin-1 (IL-1) was shown to stimulate in vitro colony formation in human bone marrow cultures. Day 4 myeloid cluster-forming cells (CFC), as well as early (day 7) and late (day 10) granulocyte-macrophage colony-forming units (CFU-GM) were stimulated in a dose-dependent fashion. At optimal concentrations of IL-1, the number of day 4 CFC reached 72%, the number of day 7 CFU-GM reached 32%, and the number of day 10 CFU-GM reached 80% of the respective numbers of colonies obtained by addition of crude leukocyte-conditioned medium (LCM). The IL-1-induced stimulatory effect on CFU-GM growth could be completely neutralized by a rabbit anti-IL-1 antiserum. Colony growth was abrogated by depleting the marrow cell suspensions of phagocytic cells prior to IL-1 addition. Conversely, the effect could be reintroduced by addition of marrow-derived adherent cells to bone marrow cell suspensions that had been depleted of both phagocytic and E rosetting T cells. Furthermore, media conditioned by bone marrow-derived adherent cells or by peripheral blood mononuclear phagocytes in the presence but not in the absence of IL-1, stimulated in vitro colony growth of phagocyte-depleted bone marrow cell suspensions. These results indicate that IL-1 induces release of granulocyte-macrophage colony-stimulating activity (GM-CSA) from human mononuclear phagocytes.

scholarly journals Brief Report: The Di Guglielmo Syndrome: Studies in Hemoglobin Synthesis

Blood ◽  
1967 ◽  
Vol 29 (4) ◽  
pp. 550-553 ◽  
Author(s):  
THOMAS F. NECHELES ◽  
WILLIAM DAMESHEK
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cell ◽  
Clinical Course ◽  
Marrow Cell ◽  
Cell Suspensions ◽  
In Vitro Synthesis ◽  
Hemoglobin Synthesis ◽  
Heme Synthesis ◽  
Globin Synthesis

Abstract The in vitro synthesis of heme and globin has been studied in bone marrow cell suspensions obtained from five patients with Di Guglielmo syndrome. In all, a defect of heme synthesis was demonstrated, but globin synthesis was greatly reduced in only two of the five; in these two, the clinical course was a rapid one.

scholarly journals INDUCTION OF A HEMOLYSIN RESPONSE IN VITRO

1970 ◽  
Vol 132 (6) ◽  
pp. 1267-1278 ◽  
Author(s):  
Klaus-Ulrich Hartmann
Keyword(s):  
Bone Marrow ◽  
Immune Response ◽  
T Cells ◽  
Cell Suspension ◽  
Cell Population ◽  
Cell Suspensions ◽  
Spleen Cells ◽  
Bone Marrow Derived Cells ◽  
Thymus Cells

The immune response to foreign erythrocytes was studied in vitro. Two subpopulations of cells were prepared. One was a population of bone marrow-derived spleen cells, taken from thymectomized, irradiated, and bone marrow-reconstituted mice; there was evidence that most of the precursors of the PFC had been present in this cell population, but few PFC developed in cultures of these cells alone in the presence of immunogenic erythrocytes. Another cell suspension was made from spleens of mice which had been irradiated and injected with thymus cells and erythrocytes; these cells were called educated T cells. The two cell suspensions together allow the formation of PFC in the presence of the erythrocytes which were used to educate the T cells, but not in the presence of noncross-reacting erythrocytes. If bone marrow-derived cells and T cells were kept in culture together with two different species of erythrocytes, and if one of the erythrocytes had been used to educate the T cells, then PFC against each of the erythrocytes could be detected.

scholarly journals Colony formation in vitro by leukemic cells in acute lymphoblastic leukemia (ALL)

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 712-718 ◽  
Author(s):  
SD Smith ◽  
EM Uyeki ◽  
JT Lowman
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Bone Marrow Cells ◽  
Lymphoblastic Leukemia ◽  
Lymphoid Cells ◽  
Assay System ◽  
Leukemic Cells ◽  
Specific Cell ◽  
Specific Cell Surface

An assay system in vitro for the growth of malignant lymphoblastic colony-forming cells (CFC) was established. Growth of malignant myeloblastic CFC has been previously reported, but this is the first report of growth of malignant lymphoblastic CFC. Established assay systems in vitro have been very helpful in elucidating the control of growth and differentiation of both normal and malignant bone marrow cells. Lymphoblastic CFC were grown from the bone marrow aspirates of 20 children with acute lymphoblastic leukemia. Growth of these colonies was established on an agar assay system and maintained in the relative hypoxia (7% oxygen) of a Stulberg chamber. The criteria for malignancy of these colonies was based upon cellular cytochemical staining characteristics, the presence of specific cell surface markers, and the ability of these lymphoid cells to grow without the addition of a lymphoid mitogen. With this technique, specific nutritional requirements and drug sensitivities can be established in vitro, and these data may permit tailoring of individual antileukemic therapy.

Reactive Oxygen Species Target the Degradation of the EVI1 Oncoprotein.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 651-651
Author(s):  
David Shackelford ◽  
You Wang ◽  
Samuel Waxman ◽  
Ruibao Ren
Keyword(s):  
Reactive Oxygen Species ◽  
De Novo ◽  
Bone Marrow Cells ◽  
Reactive Oxygen ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Mouse Bone Marrow ◽  
Oxygen Species ◽  
Aberrant Expression

Abstract Aberrant expression of EVI1 has been frequently found in myeloid malignancies as well as in cancers of the ovary, lung, head and neck, cervix, and breast, and is associated with a poor patient survival. Targeted degradation of oncoproteins is an effective strategy for cancer therapy. The AML1/MDS1/EVI1 (AME) transcription factor fusion protein is a product of the human t(3;21)(q26;q22) translocation found as a secondary mutation in some cases of CML during the blast phase (CML-BP) and in patients with de novo and therapy-related myelodysplastic syndrome (MDS) and acute myelogenous leukemia (AML). Expression of AME in mouse bone marrow cells by retroviral transduction impairs hematopoiesis and eventually induces an acute myeloid leukemia (AML)-like disease in mice. Arsenic Trioxide (ATO) has been found to be an effective treatment for patients suffering from acute promyelocytic leukemia (APL). This is, at least in part, mediated by degradation of the PML/RARα oncoprotein that is associated with over 90% of APL. We have recently shown that ATO used at therapeutic levels also degrades AME. The ATO treatment induces differentiation and apoptosis in AME leukemic cells in vitro and causes decrease in peripheral leukemic cells and splenomegaly in vivo. ATO appears to target AME at both the EVI1 and MDS moieties of the protein for degradation via the ubiquitin-proteasome pathway and proteasome-independent mechanism, respectively. To investigate the mechanism of ATO induced degradation of EVI1 oncoproteins, we examined the effect of reactive oxygen species (ROS), on EVII expression, one of the downstream effectors of ATO-induced apoptosis. We found that EVI1 degradation correlates with the amount of ROS generated in cells. EVI1 can also be targeted for degradation by doxorubicin, a chemotherapeutic agent that is effective in treating AML and ovarian cancer and a strong ROS inducer. Interestingly, the antioxidant N-acetyl cysteine (NAC) abrogates the degradation of the EVI1 protein in the presence of doxorubicin. These results demonstrate that EVI1 can be targeted for degradation by ROS. ROS inducing agents could be used as a part of targeted therapy for EVI1-positive malignancies.

Chronic Myelogenous Leukemia Cells Contribute to a Bone Marrow-Stroma in In Vivo NOD/SCID Mouse System.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2191-2191
Author(s):  
Ryosuke Shirasaki ◽  
Haruko Tashiro ◽  
Yoko Oka ◽  
Toshihiko Sugao ◽  
Nobu Akiyama ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Scid Mouse ◽  
Mononuclear Cells ◽  
Bone Marrow Cells ◽  
Myelogenous Leukemia ◽  
Bone Marrow Stroma ◽  
Marrow Stroma ◽  
Mouse System

Abstract Abstract 2191 Poster Board II-168 Aims: The stroma-forming cells in a bone marrow are derived from hematopoietic stem cells. We reported previously that non-adherent leukemia blast cells converted into myofibroblasts to create a microenvironment for proliferation of leukemia blasts in vitro. In this report we demonstrate that with severe combined immunodeficiency (SCID) mouse system chronic myelogenous leukemia (CML) cells are also differentiated into myofibroblasts to contribute to a bone marrow-stroma in vivo. Materials and Methods: Bone marrow cells were collected from informed CML patients, from which mononuclear cells were separated with density-gradient sedimentation method. After discarded an adherent cell-fraction, non-adherent mononuclear cells were injected to the priory 2.5 Gray-irradiated non-obese diabetes (NOD)/SCID mice intravenously. For the inactivation of NK cells, anti-Asialo GM1 antibody was injected intra-peritoneally prior to the transplantation, and on each 11th day thereafter. Blood was collected to monitor Bcr-Abl transcript, and mice were sacrificed after chimeric mRNA was demonstrated. Bone marrow cells were obtained, and sorted with anti-human CD133 antibody and -CD106 to select CML-derived human stromal myofibroblasts referred to the in vitro data. The isolated positive fraction was further cultured, and the biological and the molecular characteristics were analyzed. Results and Discussion: When non-adherent CML cells were transplanted to NOD/SCID mice, CML cells were engrafted after 2 months. In the murine bone marrow human stromal cells were identified, in which BCR and ABL gene was fused with FISH analysis. When the parental CML cells were cultured on the CML-derived myofibroblasts, CML cells grew extensively in a vascular endothelial growth factor-A-dependent fashion. These results indicate that CML cells can create their own microenvironment for proliferation in vivo. Disclosures: No relevant conflicts of interest to declare.

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