scholarly journals Some Factors Influencing Granulocytic Colony Formation In Vitro by Human White Blood Cells

Blood ◽  
1974 ◽  
Vol 44 (4) ◽  
pp. 511-515 ◽  
Author(s):  
Wolfgang Heit ◽  
Peter Kern ◽  
Bernhard Kubanek ◽  
Hermann Heimpel
Keyword(s):  
Colony Formation ◽  
Mononuclear Cells ◽  
White Blood Cells ◽  
Soft Agar ◽  
Red Cell ◽  
Neutrophilic Granulocytes ◽  
Feeder Layers ◽  
Granulocytic Colony ◽  
Human White Blood Cells

Abstract The experiments presented in this paper were designed to characterize the colony-stimulating activity (CSA) from white blood cell feeder layers on colony formation by peripheral blood CFCs in soft agar double-layer cultures. The data confirm earlier observations that mononuclear cells are essential for CSA production but document the importance of neutrophilic granulocytes to enhance colony formation. Possible differences between colony enhancement by granulocytes and red cell hemolysate are discussed.

scholarly journals Diphenylhydantoin-induced pure red cell aplasia

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 789-794 ◽  
Author(s):  
EN Dessypris ◽  
S Redline ◽  
JW Harris ◽  
SB Krantz
Keyword(s):  
Colony Formation ◽  
Cytotoxic Effect ◽  
Autologous Blood ◽  
Pure Red Cell Aplasia ◽  
Red Cell ◽  
Erythroid Progenitors ◽  
Red Cell Aplasia ◽  
Serum Igg ◽  
Allogeneic Marrow

Abstract The pathogenesis of diphenylhydantoin-induced pure red cell aplasia was investigated in the case of a 32-year-old man who developed pure red cell aplasia while he was under treatment with diphenylhydantoin. The patient's serum IgG purified from serum drawn at the time of diagnosis suppressed normal allogeneic marrow colony-forming (CFU-E) and burst- forming (BFU-E) and autologous blood BFU-E growth in vitro only in the presence of diphenylhydantoin. This IgG-diphenylhydantoin complex had no effect on CFU-GM growth in vitro. Normal IgG or patient's IgG purified from serum drawn after the remission of red cell aplasia had no effect on erythroid colony formation in vitro in the presence of diphenylhydantoin. The IgG-diphenylhydantoin complex exerted no direct cytotoxic effect on normal marrow erythroblasts, CFU-E, and BFU-E, nor did it interfere with the action of erythropoietin on marrow erythroblasts. These studies suggest that diphenylhydantoin-induced red cell aplasia is immunologically mediated through an IgG inhibitor, which requires the presence of the drug to suppress erythroid colony formation in vitro. This inhibitor seems to exert its effect on erythroid progenitors at or beyond the stage of differentiation of CFU- E, but not on erythroblasts.

scholarly journals THE EFFECT OF SKIN HOMOGRAFT REJECTION ON RECIPIENT AND DONOR MIXED LEUKOCYTE CULTURES

1965 ◽  
Vol 122 (4) ◽  
pp. 651-664 ◽  
Author(s):  
Joost J. Oppenheim ◽  
Jacqueline Whang ◽  
Emil Frei
Keyword(s):  
Mononuclear Cells ◽  
Tritiated Thymidine ◽  
White Blood Cells ◽  
Calf Serum ◽  
Lymphocyte Transformation ◽  
Humoral Factors ◽  
Unstimulated Control ◽  
Graft Donor

The lymphocyte proliferation in repeatedly studied mixed leukocyte cultures of peripheral white blood cells from a skin graft donor and 2 recipients was significantly increased at the time of graft rejection. This was determined from the increased proportions of mononuclear cells labeling with tritiated thymidine, increased mitotic indices, and the appearance of increased numbers of transformed lymphocytes after rejection of 1st and 2nd skin grafts. The temporarily enhanced response occurred sooner and was of shorter duration after the second than after the first graft, but was quantitatively similar each time. The cell proliferation in the mixed leukocyte cultures of the two recipients was similarly affected by the homograft rejections. The cultures containing three cell populations usually manifested a greater lymphocyte response than corresponding cultures of leukocytes from only two unrelated subjects. An increase in the ratio of female recipient to male graft donor metaphases in the cultures at the time of enhanced lymphocyte transformation indicated that proliferation of the graft recipient lymphocytes was responsible for the above findings. Unmixed, unstimulated control cultures grown in autologous, the other subjects plasma, or heterologous calf serum failed to support significant lymphocyte transformation. The role of humoral factors and relationship of the in vitro cellular responses to the in vivo homograft reaction are discussed.

Pioglitazone Inhibits the Growth of Human Leukemic Cell Lines and Primary Leukemic Cells in Vitro.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4493-4493 ◽  
Author(s):  
Yoshihiro Hatta ◽  
Minoru Saiki ◽  
Yuko Enomoto ◽  
Shin Aizawa ◽  
Umihiko Sawada ◽  
...  
Keyword(s):  
Cell Lines ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Hematopoietic Cells ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Peroxisome Proliferator ◽  
Leukemic Cell Lines ◽  
Ppar Γ

Abstract Troglitazone and pioglitazone are one of thiazolidinediones that are high affinity ligand for the nuclear receptor called peroxisome proliferator-activated receptor gamma (PPAR-γ). Troglitazone is a potent inhibitor of clonogenic growth of acute myeloid leukemia cells when combined with a retinoid. However, the effect of pioglitazone to neoplastic cells and normal hematopoietic cells has not been studied yet. Adult T-cell leukemia (ATL), prevalent in western Japan, is a highly aggressive malignancy of mature T lymphocyte. Therefore, we studied antitumor effect of pioglitazone against leukemic cells including ATL as well as normal hematopoietic cells. With 300 μM of pioglitazone, colony formation of ATL cell lines (MT1, MT2, F6T, OKM3T, and Su9T01) was completely inhibited. Colony formation of HUT102, another ATL cell line, was 12 % compared to untreated control. Clonogenic cells of other leukemic cell lines (K562, HL60, U937, HEL, CEM, and NALM1) was also inhibited to 0–30% of control. Colony formation of primary leukemic cells from 5 AML patients was decreased to 15 %. However, normal hematopoietic cells were weakly inhibited with 300 μM pioglitazone; 77 % of CFU-GM, 70 % of CFU-E, and 33 % of BFU-E survived. Cell cycle analysis showed that pioglitazone decreased the ratio of G2/M phase in HL60 cells, suggesting the inhibition of cell division. By Western blotting, PPAR-γ protein level was similar in all leukemic cells and normal bone marrow mononuclear cells. Taken together, pioglitazone effectively eliminate leukemic cells and could be used as an antitumor agent in vivo.

scholarly journals Platelet factor 4 inhibits human megakaryocytopoiesis in vitro

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1234-1239 ◽  
Author(s):  
ZC Han ◽  
L Sensebe ◽  
JF Abgrall ◽  
J Briere
Keyword(s):  
Progenitor Cells ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Platelet Factor 4 ◽  
Immunoperoxidase Staining ◽  
High Concentration ◽  
Plasma Clot ◽  
Platelet Factor ◽  
Colony Forming Unit

Platelet factor 4 (PF4) is a multifunctional protein specific to platelets, synthesized in megakaryocytes and stored in alpha granules. This report of our work shows that PF4 potently inhibits human megakaryocyte colony formation in vitro. Colony formation by megakaryocyte progenitor cells from normal bone marrows was studied using the plasma clot culture system and indirect immunoperoxidase staining. Nonadherent mononuclear cells were co-cultured with various concentrations (0 to 20 micrograms/mL) of highly purified human PF4. Statistically significant inhibition of three classes of megakaryocyte progenitor cells, the mixed colony forming unit-megakaryocytes (mCFU- MK), the burst forming unit-megakaryocytes (BFU-MK), and the colony forming unit-megakaryocytes (CFU-MK), was seen at a PF4 concentration of 2.5 micrograms/mL or greater. PF4 had no effect on erythroid (BFU-E) and granulocyte/macrophage (CFU-GM) colony formation except at high concentration (5 micrograms/mL for BFU-E and 10 micrograms/mL for CFU- GM). When a concentration of 5 micrograms/mL PF4 was added at various time points during marrow culture, a reduction of megakaryocyte colony formation also occurred. In the presence of PF4 2.5 micrograms or 5 micrograms/mL, the percentage of mature type of colonies was found to be decreased compared with cultures with no added PF4. These data demonstrate that PF4 inhibits both proliferation and maturation of megakaryocyte progenitor cells in vitro and suggest that PF4 may play a role in autoregulating human megakaryocytopoiesis.

scholarly journals Enhancement of in vitro beta-thalassemic and normal hematopoiesis by a noncytotoxic monoclonal antibody, 9.1C3: evidence for negative regulation of hematopoiesis by monocytes and natural killer cells

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1124-1133
Author(s):  
G Kannourakis ◽  
GR Johnson ◽  
CG Begley ◽  
JA Werkmeister ◽  
GF Burns
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Natural Killer Cells ◽  
Natural Killer ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Regulatory Control ◽  
Beta Thalassemia ◽  
Killer Cells

The enhancement of in vitro human hematopoiesis by the addition of a noncytotoxic monoclonal antibody, 9.1C3, is described. Enhancement of all aspects of in vitro hematopoiesis was observed on addition of 9.1C3 antibody to cultures of mononuclear cells from normal bone marrow, cord blood, and peripheral blood from beta-thalassemia major patients. In cultures with no exogenous colony-stimulating factor (CSF), the addition of 9.1C3 resulted in a two- to eightfold increase in nonerythroid colony formation. Similarly, for cultures maximally stimulated with CSF, the addition of 9.1C3 antibody resulted in a one- to fourfold increase in colony formation. These effects were abrogated by the removal of either adherent, Leu-M3+ or Leu-7+ cells. Colony- forming cells were shown to be present among the 9.1C3-negative cells when mononuclear cells were sorted by flow cytometry. Media conditioned in the presence of 9.1C3 and mononuclear cells were able to enhance colony formation in vitro for normal nonadherent bone marrow cells beyond that achieved with supramaximal amounts of human placental- conditioned medium and erythropoietin. The data suggest that natural killer cells interact with monocytes to exert a negative regulatory control on in vitro granulopoiesis and erythropoiesis. Consequently, the number of progenitor and multipotential cells in cultures of unfractionated cell populations may be greatly underestimated.

scholarly journals Effect of peripheral blood mononuclear cells from aplastic anemia patients on the granulocyte-macrophage and erythroid colony formation in samples from normal human bone marrow in vitro--a cooperative work

Blood ◽  
1980 ◽  
Vol 55 (6) ◽  
pp. 937-943
Author(s):  
F Takaku ◽  
T Suda ◽  
H Mizoguchi ◽  
Y Miura ◽  
H Uchino ◽  
...  
Keyword(s):  
Aplastic Anemia ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Suppressive Effect ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Normal Human

Thirty-six patients with aplastic anemia and three with aplastic anemia- PNH syndrome in eight institutes were studied for the presence of peripheral blood mononuclear cells having a suppressive effect on granulocyte-macrophage (G-M) and erythroid colony formation using a uniform protocol. In 11 cases of 29 (38%) and 9 cases of 29 (31%), the presence of mononuclear cells with a suppressive effect on G-M and erythroid colony formation, respectively, was demonstrated. However, the presence of mononuclear cells suppressive both to G-M and erythroid colony formation was demonstrated only in 3 of 19 cases (16%).

scholarly journals Inhibition of CFU-NM and CFU-EOS by mature granulocytes

Blood ◽  
1984 ◽  
Vol 63 (2) ◽  
pp. 376-379
Author(s):  
BH Bjornson ◽  
SH Pincus ◽  
AM DiNapoli ◽  
JF Desforges
Keyword(s):  
Mononuclear Cells ◽  
Feedback Inhibition ◽  
Feeder Layer ◽  
Intact Cells ◽  
Normal Peripheral Blood ◽  
Inhibition Of Growth ◽  
Feeder Layers ◽  
Eosinophil Colonies ◽  
Differential Control

Approximately half of the colony-forming units-culture (CFU-C) from normal peripheral blood are eosinophilic. The purpose of our study was to determine: (1) whether progenitor cells committed to eosinophil or neutrophil maturation would be differentially affected by feedback inhibition, and (2) whether mature eosinophils added to the feeder layers of the culture would inhibit the proliferation of CFU-C in a manner similar to that described for neutrophils. Concentrated eosinophils and neutrophils, obtained by separation on a metrizamide gradient, were added to feeder layers containing either 10(6) autologous whole mononuclear cells (WMNC) or 0.1 ml of leukocyte conditioned media (LCM). The average number of colonies was 123/10(6) nonadherent cells (NAC) cultured. When neutrophils or eosinophils were added to the WMNC feeder layer, the percent inhibition of growth was 40.2% +/- 1.6% (mean +/- SEM) and 42.3% +/- 5.4%, respectively, but the ratio of neutrophil to eosinophil colonies remained constant. No effect was seen when neutrophils or eosinophils were added to an LCM feeder layer. Thus, it appears that the differential control of neutrophil versus eosinophil production in vitro is not regulated through feedback inhibition by mature granulocytes. In addition, these studies suggest that eosinophils, as well as neutrophils, cause inhibition of CFU-C growth when intact cells are the source of colony-stimulating factor (CSF).

scholarly journals Interrelationship between mitosis and endomitosis in cultures of human megakaryocyte progenitor cells [published erratum appears in Blood 1987 May;69(5):1547]

Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 486-492 ◽  
Author(s):  
M Arriaga ◽  
K South ◽  
JL Cohen ◽  
EM Mazur
Keyword(s):  
Colony Formation ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Geometric Mean ◽  
Cell Diameter ◽  
Prior History ◽  
Serum Concentrations ◽  
Cell Numbers ◽  
Canine Serum

Abstract Sera from dogs rendered aplastic by total-body irradiation stimulate human bone marrow megakaryocyte progenitors to form megakaryocyte colonies in plasma clot cultures. In this investigation, we evaluated the effects of varying concentrations of such sera on both the mitotic and endomitotic phases of human megakaryocyte development in vitro. When low concentrations of aplastic canine sera (2.5% to 5.0% [vol/vol]) were added to cultures of human peripheral blood mononuclear cells in place of normal AB serum, megakaryocyte colony formation was augmented fivefold, cell numbers per colony increased approximately 2.5- fold, and the geometric mean megakaryocyte ploidy almost doubled. Further increasing the aplastic canine serum concentration from 10% to 30% (vol/vol) stimulated no additional colony formation. However, there was a further augmentation of cell numbers per colony associated with a progressive decrease in the mean megakaryocyte ploidy. Megakaryocyte cultures were harvested after 7, 12, 15, and 19 days of incubation, and these demonstrated that the lower mean ploidy values found at the higher concentrations of aplastic canine serum did not result from delayed endoreduplication. At all aplastic serum concentrations evaluated, there existed a strong correlation between nuclear ploidy and cell diameter. We conclude that both the mitotic and endomitotic events in human megakaryocytopoiesis may be influenced by a factor or factors present in aplastic canine serum. At lower in vitro concentrations, such sera stimulate both mitosis and endomitosis, which promotes the development of megakaryocyte colonies composed of larger cells with a higher mean ploidy. With increasing aplastic serum concentrations, colony formation plateaus and mitosis is favored over endomitosis. This results in colonies composed of more numerous but smaller megakaryocytes with a lower mean ploidy. Our data suggest that the size and extent of polyploidization that can be achieved by a developing megakaryocyte may be influenced by the mitotic prior history of its immediate precursor cell.

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