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 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

Abstract 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.

Thrombopoietin-Induced Stimulation of Megakaryocyte- Enriched Bone Marrow Cultures

1979 ◽  
Author(s):  
K. L. Kellar ◽  
B. L. Evatt ◽  
C. R. McGrath ◽  
R. B. Ramsey
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Bone Marrow Cells ◽  
Rabbit Plasma ◽  
Bone Marrow Cultures ◽  
Plasma Fractions ◽  
Marrow Cultures ◽  
Stimulation Of

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.

scholarly journals Granulopoietic effects of human bone marrow fibroblastic cells and abnormalities in the "granulopoietic microenvironment"

Blood ◽  
1981 ◽  
Vol 58 (3) ◽  
pp. 557-564 ◽  
Author(s):  
BR Greenberg ◽  
FZD Wilson ◽  
L Woo
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Myeloproliferative Disorders ◽  
Conditioned Media ◽  
Myelogenous Leukemia ◽  
Acute Myelogenous ◽  
Colony Number ◽  
Fibroblastic Cells

Abstract The in vitro granulopoietic effects of adherent bone marrow fibroblastic cells (FC) were studied in normal humans and in patients with acute myelogenous leukemia (AML) and myeloproliferative disorders (MPD). To determine their influence on granulopoiesis, we established FC in liquid-phase cultures, overlaid the adherent FC with normal bone marrow cells in agar, and subsequently measured the growth of CFU-C. When using target marrows containing few spontaneous colonies, increased numbers of CFU-C were found above the FC obtained from normals. No growth greater than controls was observed in those areas lacking FC. If target marrows contained large numbers of spontaneous CFU-C, actual inhibition of colony formation was produced by FC co- incubation. In contrast to normals, FC obtained from untreated AML and MPD patients typically failed to enhance granulopoiesis. Regardless of source, FC were not synergistic with the effects of placenta- conditioned media (typically being inhibitory) for colony number, but were synergistic for colony size. Conditioned media obtained from FC cultures did not enhance colony formation and actually inhibited spontaneous colony formation. Thus, microenvironmental abnormalities in interactions between “stromal cells” and hematopoietic progenitors may be important in the pathogenesis and clinical expression of hematopoietic malignancies in humans.

scholarly journals Granulopoietic effects of human bone marrow fibroblastic cells and abnormalities in the "granulopoietic microenvironment"

Blood ◽  
1981 ◽  
Vol 58 (3) ◽  
pp. 557-564
Author(s):  
BR Greenberg ◽  
FZD Wilson ◽  
L Woo
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Myeloproliferative Disorders ◽  
Conditioned Media ◽  
Myelogenous Leukemia ◽  
Acute Myelogenous ◽  
Colony Number ◽  
Fibroblastic Cells

The in vitro granulopoietic effects of adherent bone marrow fibroblastic cells (FC) were studied in normal humans and in patients with acute myelogenous leukemia (AML) and myeloproliferative disorders (MPD). To determine their influence on granulopoiesis, we established FC in liquid-phase cultures, overlaid the adherent FC with normal bone marrow cells in agar, and subsequently measured the growth of CFU-C. When using target marrows containing few spontaneous colonies, increased numbers of CFU-C were found above the FC obtained from normals. No growth greater than controls was observed in those areas lacking FC. If target marrows contained large numbers of spontaneous CFU-C, actual inhibition of colony formation was produced by FC co- incubation. In contrast to normals, FC obtained from untreated AML and MPD patients typically failed to enhance granulopoiesis. Regardless of source, FC were not synergistic with the effects of placenta- conditioned media (typically being inhibitory) for colony number, but were synergistic for colony size. Conditioned media obtained from FC cultures did not enhance colony formation and actually inhibited spontaneous colony formation. Thus, microenvironmental abnormalities in interactions between “stromal cells” and hematopoietic progenitors may be important in the pathogenesis and clinical expression of hematopoietic malignancies in humans.

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

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1621-1621
Author(s):  
Haruko Tashiro ◽  
Ryosuke Shirasaki ◽  
Yoko Oka ◽  
Toshihiko Sugao ◽  
Nobu Akiyama ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Scid Mouse ◽  
Mononuclear Cells ◽  
Bone Marrow Cells ◽  
Myelogenous Leukemia ◽  
Acute Myelogenous ◽  
Blast Cells ◽  
Marrow Cells

Abstract Abstract 1621 Poster Board I-647 Aims: Cancer stem cell theory has been developed, and whose precise characteristics have been reported. However, there have been no reports on the differentiation of cancer stem cells into the environmental stromal cells. We reported previously that non-adherent acute myelogenous leukemia (AML) cells were differentiated into myofibroblasts to create a microenvironment for proliferation of AML blasts in vitro. In this report we demonstrate that with severe combined immunodeficiency (SCID) mouse system AML blast cells also convert to myofibroblasts to form stroma in vivo. Materials and Methods Bone marrow cells were collected from informed AML (M2) patients who had chromosomal translocation of RUNX1 and ETO, from which mononuclear cells were separated with density-gradient sedimentation method. After discarded an adherent cell-fraction, the non-adherent mononuclear cells were injected to the 3.0 Gray-irradiated non-obese diabetes (NOD)/SCID mouse 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 Runx1 and ETO fusion transcript, and mice were sacrificed after chimeric mRNA was observed. Bone marrow cells were obtained, and sorted with anti-human CD133 antibody and -CD106 to select AML-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 AML (M2) blast cells were transplanted to NOD/SCID mice, cells were engrafted after 10 weeks. In murine bone marrow cells human stromal cells were identified, in which RUNX1 and ETO gene was fused with FISH analysis. When the parental AML blast cells were cultured on the expanded AML-derived myofibroblasts, AML cells grew extensively. These results indicate that AML cells can create their own microenvironment for proliferation in vivo. Disclosures No relevant conflicts of interest to declare.

scholarly journals Interleukin-3 is significantly more effective than other colony- stimulating factors in long-term maintenance of human bone marrow- derived colony-forming cells in vitro

Blood ◽  
1989 ◽  
Vol 73 (7) ◽  
pp. 1836-1841 ◽  
Author(s):  
M Kobayashi ◽  
BH Van Leeuwen ◽  
S Elsbury ◽  
ME Martinson ◽  
IG Young ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cultured Cells ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Factor G ◽  
Marrow Cells

Abstract Human bone marrow cells cultured for 21 days in the presence of recombinant human interleukin-3 (IL-3) produced up to 28 times more colony-forming cells (CFC) than could be obtained from cultures stimulated with granulocyte colony stimulating factor (G-CSF) or granulocyte-macrophage CSF (GM-CSF). IL-3-cultured cells retained a multipotent response to IL-3 in colony assays but were restricted to formation of granulocyte colonies in G-CSF and granulocyte or macrophage colonies in GM-CSF. Culture of bone marrow cells in IL-3 also led to accumulation of large numbers of eosinophils and basophils. These data contrast with the effects of G-CSF, GM-CSF, and IL-3 in seven-day cultures. Here both GM-CSF and IL-3 amplified total CFC that had similar multipotential colony-forming capability in either factor. G-CSF, on the other hand, depleted IL-3-responsive colony-forming cells dramatically, apparently by causing these cells to mature into granulocytes. The data suggest that a large proportion of IL-3- responsive cells in human bone marrow express receptors for G-CSF and can respond to this factor, the majority becoming neutrophils. Furthermore, the CFC maintained for 21 days in IL-3 may be a functionally distinct population from that produced after seven days culture of bone marrow cells in either IL-3 or GM-CSF.

scholarly journals Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures.

1989 ◽  
Vol 9 (9) ◽  
pp. 3973-3981 ◽  
Author(s):  
G V Borzillo ◽  
C J Sherr
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Chain Gene ◽  
Mouse Bone Marrow ◽  
Feeder Layers ◽  
Bone Marrow Cultures ◽  
B Lymphoid ◽  
Marrow Cultures

Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B-lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage.

scholarly journals Heme-stress activated NRF2 skews fate trajectories of bone marrow cells from dendritic cells towards red pulp-like macrophages in hemolytic anemia

2022 ◽  
Author(s):  
Florence Vallelian ◽  
Raphael M. Buzzi ◽  
Marc Pfefferlé ◽  
Ayla Yalamanoglu ◽  
Irina L. Dubach ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Dendritic Cell ◽  
Sickle Cell Disease ◽  
Bone Marrow Cells ◽  
Cell Disease ◽  
Secondary Immunodeficiency ◽  
Gm Csf ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

AbstractHeme is an erythrocyte-derived toxin that drives disease progression in hemolytic anemias, such as sickle cell disease. During hemolysis, specialized bone marrow-derived macrophages with a high heme-metabolism capacity orchestrate disease adaptation by removing damaged erythrocytes and heme-protein complexes from the blood and supporting iron recycling for erythropoiesis. Since chronic heme-stress is noxious for macrophages, erythrophagocytes in the spleen are continuously replenished from bone marrow-derived progenitors. Here, we hypothesized that adaptation to heme stress progressively shifts differentiation trajectories of bone marrow progenitors to expand the capacity of heme-handling monocyte-derived macrophages at the expense of the homeostatic generation of dendritic cells, which emerge from shared myeloid precursors. This heme-induced redirection of differentiation trajectories may contribute to hemolysis-induced secondary immunodeficiency. We performed single-cell RNA-sequencing with directional RNA velocity analysis of GM-CSF-supplemented mouse bone marrow cultures to assess myeloid differentiation under heme stress. We found that heme-activated NRF2 signaling shifted the differentiation of bone marrow cells towards antioxidant, iron-recycling macrophages, suppressing the generation of dendritic cells in heme-exposed bone marrow cultures. Heme eliminated the capacity of GM-CSF-supplemented bone marrow cultures to activate antigen-specific CD4 T cells. The generation of functionally competent dendritic cells was restored by NRF2 loss. The heme-induced phenotype of macrophage expansion with concurrent dendritic cell depletion was reproduced in hemolytic mice with sickle cell disease and spherocytosis and associated with reduced dendritic cell functions in the spleen. Our data provide a novel mechanistic underpinning of hemolytic stress as a driver of hyposplenism-related secondary immunodeficiency.

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