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Results for “Bone Marrow Cells”

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At least 19 recordsLinked to original sources

PubMed1989

The effects of demineralized bone matrix and direct current on an "in vivo" culture of bone marrow cells.

Bone marrow cells (BMCs) from rabbit femora and tibiae were grown in diffusion chambers implanted in rabbit muscle. At 42 days 80% of the BMC chambers exhibited cartilage formation within them. Demineralized bone matrix added to the marrow cell suspension in the chamber accelerated the appearance and increased the number of chambers with cartilage. Mineralization of the cartilage also occurred earlier in the chambers with bone matrix. In a second experiment, a 5-microA direct current cathode in the bone marrow chamber increased the number of chambers containing cartilage from 50 to 80% at day 25. Mineralization also occurred earlier in the chambers with direct current.

Animals↗

PubMed1988

Chromosome breaks and fragile sites in leukemic bone marrow cells.

Bone marrow cells from leukemic and nonleukemic patients were examined for chromosome breakage in cultures treated with fluorodeoxyuridine (FUdR) and FUdR plus caffeine. The results indicate that the leukemic cells have more chromosome breakage than the nonleukemic cells when thymidylic synthetase is inhibited by FUdR. Addition of caffeine did not enhance this chromosome breakage. These findings of enhanced breakage by FUdR exposure in vitro, nevertheless, may suggest that leukemic cells in general are more susceptible to breakage than normal cells, thereby predisposing the former to secondary chromosome rearrangements.

Adolescent↗

PubMed1988

[In vitro manipulation of bone marrow cells for bone marrow transplantation].

Recently, in vitro manipulations of bone marrow cells have been developed for the prevention of relapse in autologous bone marrow transplantation (BMT) and for the prevention of graft versus host disease in allogeneic BMT. Two methods were mainly employed clinically for the depletion of cells from transplanted bone marrow cells. Firstly, bone marrow cells were treated in vitro with monoclonal antibodies reactive to leukemia cells or T cells using complement, immunotoxin or magnetic bead. Secondly, bone marrow cells were incubated with cancer drugs such as 4-HC or mafosfamide. Our results and other reports using in vitro purging of leukemia or lymphoma cells suggest that these autologous BMTs are effective modes of cancer therapy in patients with hematological malignancies.

Acute Disease↗

PubMed1979

Induction of erythropoietin responsiveness in vitro by a distinct population of bone marrow cells.

Bone marrow contains a small population of primitive erythroid progenitor cells which can be detected by their capacity to form large numbers of erythroid progeny in viscous cultures containing erythropoietin (EP). These cells have been termed erythroid 'burst-forming units' (BFUe). The present study demonstrates that expression of the erythroid differentiation potential of BFUe requires the presence of an activity additional to EP. This activity has been designated as BFA (burst feeder activity). It is shown that the number of BFUe detected and their apparent sensitivity to EP are directly related to the BFA concentration of the cultures. BFA was found to be associated with a population of bone marrow cells of high buoyant density and small volume, which are sensitive to irradiation. The radiation dose-effect curve provided strong evidence that bone marrow BFA is independent of cell proliferation; this was supported by showing that BFA is unaffected by in vivo treatment with hydroxyurea. The findings are compatible with a two-step regulation model for erythroid differentiation in which BFA-induced progeny of BFUe acquire sensitivity to EP.

Animals↗

PubMed1979

Dilution techniques for optimum recovery of cryopreserved bone marrow cells.

Bone marrow granulocyte progenitor cells (CFU-C) were assayed in methyl-cellulose prior to cryopreservation in Dimethylsulfoxide (DMSO) and after thawing and diluting the DMSO. The time of dilution from 10% to 1% DMSO and the temperature of the sample and diluting media were studied. Compared with samples diluted at 0 degrees-4 degrees C, samples which were diluted at 24 degrees C were more viable by Trypan Blue exclusion (p less than .01) and had greater CFU-C growth in vitro (p less than .01). There was no advantage to prolonging dilution time from 10 minutes at a constant rate to 40 minutes using stepwise technique. Recovery of CFU-C at 24 degrees C ranged from 40% to 114% with a mean +/- S.D. of 67% +/- 19.5%. There was evidence that clonogenic cells were selectively preserved under the conditions described.

Bone Marrow Cells↗

PubMed1985

Characterization of cytotoxic cells generated from in vitro cultures of murine bone marrow cells.

Bone marrow cells cultured for 5-6 days generate cytotoxic activity against a number of natural killer (NK)-susceptible tumor cells. In this study, these bone marrow cytotoxic cells were compared to cells with NK activity obtained either from spleen cells activated in vitro with interferon (IFN-alpha/beta) or mitogen or from peritoneal exudate cells (PEC) obtained 4 days after bacillus Calmette-Guerin (BCG) infection. Splenic and PEC cytotoxic cells were shown to be Thy 1.2+, NK 1.1+, Asialo GM+1, Lyt 1.2-, Lyt 2.2-. In contrast, bone marrow cytotoxic cells were Thy 1.2+, NK 1.1-, Lyt 1.2-, Lyt 2.2- and expressed low levels of Asialo GM1 antigen (Asialo GM +/- 1). Precursor cells for bone marrow cytotoxic activity were shown to be Thy 1.2-, NK 1.1-, Lyt 1.2-, Lyt 2.2- but also expressed low levels of Asialo GM1 antigen (Asialo GM +/- 1). Cytotoxic activity for both bone marrow and spleen cells peaked in the low-density fractions of discontinuous Percoll density gradients. The cytotoxic activity of these bone marrow cells was augmented by pretreatment with IFN (-alpha/beta, -gamma) or soluble factors (IFN free) from activated EL-4 thymoma cells. Surprisingly, the ability of bone marrow cells to generate high levels of cytotoxic activity following in vitro culture appeared to be associated primarily with mice which were of the H-2b haplotype.

Animals↗

PubMed1991

Reactivity of the workshop monoclonal antibodies with ovine bone marrow cells and bone marrow-derived monocyte/macrophage and mast cell lines.

The workshop monoclonal antibodies were tested by flow cytometry for reactivity against: (1) ovine bone marrow cells, (2) cultured bone marrow-derived monocyte/macrophage cell lines and (3) cultured bone marrow-derived mast cell lines. Both single and two-colour immunofluorescence tests were performed. The results of these analyses are presented and discussed.

Animals↗

PubMed1986

Chromosome aberrations in rat liver cells and bone marrow cells following treatment in vivo with mitomycin C.

The clastogenic potential of mitomycin C (MMC) was studied in rat liver cells and bone marrow cells. Male Sprague-Dawley rats were partially hepatectomized and treated with a single i.p. dose of MMC (3.5 mg/kg body weight) 7 or 24 h after the operation. Non-hepatectomized rats were also treated with the same dose of MMC 7 or 24 h after the mutagenic treatment; liver and bone marrow cells were isolated from hepatectomized rats (31 h after the operation) and bone marrow cells only from non-hepatectomized animals. The results show that, if MMC was administered 7 h before the isolation of cells, the induction was more efficient in liver cells than in bone marrow cells. At this sampling time, there was no consistent difference between the frequencies observed in bone marrow cells from hepatectomized and non-hepatectomized rats. An increase was observed in both tissues 24 h after the mutagenic treatment. At this sampling time, the effect was significantly higher in the bone marrow cells from non-hepatectomized animals than in the liver cells and bone marrow cells from hepatectomized animals. No significant difference between the two cell types from hepatectomized rats was observed. Different factors related to the cytostatic properties of MMC and/or to cell kinetics in the two cell types, probably affected by the surgical operation, may account for the differences in the yield of chromosomal damage.

Animals↗

PubMed1991

Characterization and regulation of RB6-8C5 antigen expression on murine bone marrow cells.

Murine bone marrow cells expressing the cell surface Ag RB6-8C5 were identified by fluorescence-activated cell-sorting analysis using a rat IgG mAb. The fluorescent intensity of RB6-8C5 was variable on bone marrow cells. This made it possible to separate bone marrow cells into distinct subpopulations, RB6-8C5neg, RB6-8C5lo, and RB6-8C5hi cells. Morphologic analysis of the sorted populations demonstrated that the Ag was expressed on myeloid cells. The expression of RB6-8C5 increases with granulocyte maturation, whereas expression is transient on cells in the monocytic lineage. The RB6-8C5hi sorted cells were enriched for end-stage neutrophils (75%), whereas the RB6-8C5lo sorted cells contained more immature myeloid cells and myelocytes (75%). Lymphocytes and macrophages were less than 5% in any RB6-8C5+ population, whereas the erythroid precursors were RB6-8C5neg. The colony forming unit culture (CFU-C) (greater than 90%) were found in the RB6-8C5neg and RB6-8C5lo populations, and all the CFU-granulocyte, erythroid, megakaryocyte, and macrophage (CFU-GEMM) and burst-forming units-erythroid (BFU-E) were in the RB6-8C5neg population. Granulocyte-macrophage-CSFR (GM-CSFR) and IL-1 alpha R were expressed on RB6-8C5hi bone marrow cells, whereas no receptors could be detected on RB6-8C5neg and RB6-8C5lo cells. The expression of the RB6-8C5 Ag can be induced on RB6-8C5neg cells in liquid culture by IL-3 and granulocyte-macrophage CSF. Thus, RB6-8C5 is a myeloid differentiation Ag whose expression can be regulated by cytokines.

Animals↗

PubMed1988

Bone marrow cells from allogeneic bone marrow chimeras inhibit the generation of cytotoxic lymphocyte responses against both donor and recipient cells.

When added to a mixed lymphocyte culture, bone marrow cells suppress the generation of CTL activity against H-2 Ag shared by the BM cells and the stimulator cells. These cells have been referred to as veto cells and are thought to play a role in maintaining self-tolerance. We analyzed the H-2 specificity of the suppression expressed by the veto cells from H-2 incompatible bone marrow chimeras, because lymphocytes of such chimeras had been shown to be tolerant to both donor and recipient Ag when tested by CTL responses. We found that the bone marrow cells of such chimeras which were featured by non-T and non-B cell characteristics inhibited the generation of CTL directed against either donor or recipient Ag, but not against third-party Ag. These observations suggest that in allogeneic chimeras the veto or veto-like cells alter the inhibitory specificity exhibited in the recipient microenvironment and indicate that these cells are directly involved in the induction and maintenance of self-tolerance.

Animals↗

PubMed1987

Bone marrow cells other than stem cells seed the bone marrow after rescue transfusion of fatally irradiated mice.

In a previous publication, iodinated deoxyuridine (125IUdR) incorporation data were interpreted as indicating that spleen colony-forming units (CFU-S) in DNA synthesis preferentially seeded bone marrow. In the present studies, the CFU-S content of marrow from irradiated, bone-marrow transfused mice was directly determined. Pretreatment of the transfused cells with cytocidal tritiated thymidine resulted in an insignificant diminution in CFU-S content when compared with nontritiated thymidine pretreatment, implying that there is no preferential seeding. The 125IUdR incorporation data have been reinterpreted as being a result of the proliferation of other progenitor cells present that have seeded the bone marrow.

Animals↗

PubMed1992

Effect of enkephalins on bone marrow cells.

Mouse bone marrow cells were incubated with methionine- or leucine-enkephalin (10(-15)-10(-6) M) before seeding into soft agar cultures. In marrow samples harvested at different times, enkephalins decreased GM colony count on average by 30-40%. In individual experiments, however, the same concentration of enkephalins caused even stimulation, or at other times had effect. In view of the circadian periodicity of neuroendocrine functions and hematopoietic activity, the enkephalin effect on bone marrow cells was tested on marrow samples harvested at fixed time points (6 am, 6 pm), using enkephalin concentrations in the physiological range (10(-12)-10(-9) M). The seeding efficiency of the 6-pm cell population was on average 50% above that of the 6-am population. The 6-pm cell population was also more susceptible to the inhibitory effect of the enkephalins (35% inhibition) than the 6-am population (15% inhibition), and the variability in response was considerably reduced. With progenitor cell-enriched population, obtained by fluorescence-activated cell sorting (FACS) of 6-am bone marrow samples, in 3 out of 6 experiments Met- and Leu-enkephalin showed 30-35% inhibition of GM colony formation over a wide range of concentrations (10(-15)-10(-6)). In the other 3 experiments, suppression as well as stimulation or no alteration in colony count were observed. This variability probably reflected quality (purity) of the progenitor cell population, and may indicate that the enkephalins affected hematopoietic cells via a population of accessory cells.

Analysis of Variance↗

PubMed1975

Mechanism of action of granulopoiesis inhibiting factor (chalone). I. Evidence for a receptor protein on bone marrow cells.

Rat bone marrow cells respond to granulopoiesis inhibiting factor by a reduced incorporation of tritiated thymidine. Cells treated with low concentrations of trypsin lose their ability to respond to the factor if protein synthesis is partially inhibited by low doses of cycloheximide. Responsiveness is retained if protein synthesis is permitted after enzyme treatment. The data suggest that a protein receptor on the external surface of the target cells is required for the action of granulopoiesis inhibiting factor on bone marrow cells.

Animals↗

PubMed1978

Histological and cytological technique for the quantitation of cultured human bone-marrow cells:formation of aggregates.

A bone-marrow culture system is described that provides a simple, quantitative and rapid assessment of marrow bone cells in vitro. Aggregation of bone-marrow cells, an in vitro phenomenon, occurs within 24 hr of culture and is observed utilizing Millipore filters. Daily quantitation shows both an increase in the number and a change in the morphology of these aggregates. The maximum number of aggregates is achieved on the 2nd or 3rd day of incubation. Histologically, aggregates are composed of myeloid, mononuclear and mesenchymal fibroblastic cells. Mesenchymal cells form a matrix for apposed mononuclear and myeloid cells. Scanning electron micrographs show intimate cell contact and spreading by the marrow cells. Fluctuation of the absolute numbers of various cell types are observed. The system can be utilized for long-term culture of bone marrow.

Bone Marrow Cells↗

PubMed1987

Detection of residual murine LPC-1 myeloma cells from bone marrow cell mixture after purging by 4-hydroperoxycyclophosphamide.

Mixtures of BALB/c bone marrow (5 X 10(6) and LPC-1 myeloma (2 X 10(6) cells have been purged in vitro with 100 microM 4-hydroperoxycyclophosphamide (4HC) for 30 min at 37 degrees C. Elimination of tumor cells was assessed by monitoring newly synthesized tumor-specific IgG 2a kappa in vitro and by reinjecting the cells subcutaneously into the hindlegs of mice. Drug-treated LPC-1 cells had no detectable tumor production and did not reproduce tumors. Untreated cells regrew as solid tumors and killed the host within 3-4 weeks. Bone marrow cell suspensions purged of tumor cells were then used to reconstitute lethally pretreated mice injected 24 h earlier with 300 mg/kg cyclophosphamide (CY). Mice injected with CY alone died within 10 days. Those reconstituted with bone marrow cells or with purged bone marrow-tumor cell mixture lived longer than 7 months. Mice reconstituted with untreated bone marrow-tumor cell suspension grew tumors, had detectable tumor-specific IgG 2a kappa in their serum, and died by day 44. These studies demonstrate that the success of myeloma cell purging can be determined by monitoring newly secreted tumor protein and that 4HC successfully eliminates malignant plasma cells in vitro without impairment of normal bone marrow stem cell functions.

Animals↗

PubMed1975

[The mutagenic effect of thioTEPA in laboratory mice. IV. The influence of genotype and sex on the frequency of induced chromosome aberrations in bone marrow cells].

In bone marrow cells of eleven genotypes of mice strain and sex variations in sensitivity to cytogenetic effect of thio-TEPA were studied. Strains 101/H, C57BL/6 and A/Sn were the more sensitive and F1 (CBAXC57BL/6) and F1 (C3HX101) were the more resistant to induction of chromosome damages. The rate of induced chromosome aberrations was correlated with the level of spontaneous chromosome damages in bone marrow cells (r=+/-0,893+/-0,02). From all strains males were more mutable than females. It is recommended to use C57BL/6 strain of mice male for genetic tests of chemicals by cytogenetic methods in vivo.

Animals↗

PubMed1990

T-cell growth factor P40 promotes the proliferation of myeloid cell lines and enhances erythroid burst formation by normal murine bone marrow cells in vitro.

T-cell growth factor P40 was examined for possible effects on murine interleukin-3 (IL-3)-dependent myeloid cell lines and freshly isolated murine bone marrow cells. The results showed that P40 stimulated the proliferation of some IL-3-dependent myeloid cell lines of both early myeloid and mast cell phenotype and synergized with IL-3. P40 did not promote proliferation of fresh bone marrow cells, bone marrow enriched for early myeloid cells by 5-fluorouracil treatment, or bone marrow derived mast cells as assessed in 3H-TdR incorporation assays. P40 did not influence the growth of murine colony-forming unit granulocyte-macrophage in agar cultures, either alone or in the presence of optimal or sub-optimal concentrations of CSF-1, GM-colony-stimulating factor, or IL-3. P40 did potentiate burst-forming unit-erythroid (BFU-E) formation in the presence of erythropoietin; however, this was dependent on the cell plating density, suggesting an indirect stimulation of BFU-E by P40. The indirect nature of P40 action on BFU-E was further demonstrated in cell separation experiments and indicated that the effect was mediated by T cells. These data expand the repertoire of cells that P40 influences.

Animals↗

PubMed1977

T cell function detected in murine bone marrow cells.

Mouse bone marrow cells were fractionated by BSA discontinuous density gradient centrifugation, and a small lymphocyte rich fraction was obtained at the high density. Cells of this fraction were shown to respond in vitro to T cell mitogens and alloantigens. Furthermore, they were able to mount a graft-versus-host reaction when assessed by spleen weight assay and by the method of inhibiting erythroid cell growth by allogeneic lymphoid cells. The results indicate that these lymphocytes possess T cell functions. On the other hand they were found to carry only little theta antigen assessed by the cytotoxic test and by the absorption test. It is presumed, therefore, that the amount of theta antigen on a cell might not correlate with T cell functions, and these lymphocytes might be mature ones in the course of postthymic maturation. Hemopoietic stem cells were determined by spleen colony formation and the peak of colony-forming efficiency was seen at the low density. These observations imply that immunocompetent cells causing GVHR can be separated from hemopoietic stem cells. This procedure may be applied for prevention and reduction of GVHR in allogeneic bone marrow transplantation in human.

Animals↗