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At least 19 recordsLinked to original sources
PubMed1978

Heterogeneity of human colony-forming cells (CFU-C): differences in size, rate of colony formation, and responsiveness to colony-stimulating factor.

Human bone marrow cells have been fractionated by velocity sedimentation at unit gravity. Fractions were analyzed for cell morphology, number of nucleated cells and myeloid colony-forming cells (CFU-C's). Colony formation was assayed with the following CSF preparations: serum-free HLCM and two electrophoretically distinct CSF fractions (CSF-A and CSF-B) purified from the same source. Two distinct CFU-C populations were found. One, a rapidly sedimenting (7.2 to 8.0 mm/hr), population exhibited colonies after 7 days of culture in response to HLCM and CSF-A only. The second, a more slowly sedimenting (6.5 mm/hr) CFU-C peak, did not exhibit colonies until 11 days of culture and did so in response to all three CSF's tested. The results indicate that human bone marrow CFU-C's are heterogeneous and that the two purified CSF fractions from human lung have different CFU-C specificity.

Bone Marrow↗
PubMed1978

Monoclonal origin of B lymphocyte colony-forming cells in spleen colonies formed by multipotential hemopoietic stem cells.

Spleen colonies produced by transplanting lethally irradiated mice with either 12 day fetal liver or adult bone marrow cells were found to contain B- lymphocyte colony-forming cells (BL-CFC) . The proportion of BL-CFC positive spleen colonies did not increase substantially between 8 and 14 days after transplantation, the range being 18-45 percent. However, the absolute number of BL-CFC per spleen colony varied considerably (between 1 and 10,318), although the majority of colonies contained less than 200 BL-CFC. Irrespective of the time after transplantation, smaller spleen colonies were found to have a higher frequency of BL-CFC than larger spleen colonies. To determine the possible clonal origin of BL-CFC from spleen colony- forming unit (CFU-S), CBA mice were injected with equal numbers of CBA and CBA T(6)/T(6) fetal liver or adult bone marrow cells. Analysis of 7-15-day spleen colonies demonstrated that 90 percent were either exclusively T(6) positive or T(6) negative and approximately equal numbers ofboth colony types were observed. B-lymphocyte colonies were grown and successfully karyotyped from 19 spleen colonies. When compared with the original spleen colony karyotype the B-lymphocyte colony cells karyotype was identical in all 19 cases. In 3 of the 19 colonies analyzed a mixture of T(6) positive and T(6) negative karyotypes was present and identical proportions of the karyotypes were present in the pooled B-lymphocyte colony cells and spleen colony cells. The data indicate that the B-lymphocyte colony-forming cells detected in spleen colonies are genuine members of the hemopoietic clone derived from the initiating hemopoietic stem cell (CFU-S).

Animals↗
PubMed1976

Cellular responsiveness to stimulation in vitro: increased responsiveness to colony stimulating factor of bone marrow colony-forming cells treated with surface-active agents and cyclic 3'5' AMP.

Addition of low concentrations (10 ng/ml) of saponin or Tween 80 to stimulated cultures of normal mouse bone marrow in agar increased the number of granulocyte-macrophage colonies which developed. Addition of cyclic AMP or dibutyryl cyclic AMP in low concentration (10(-8) to 10(-10) M) also enhanced colony numbers although concentrations above 10(-5) M were inhibitory. enhancement was found when marrow cells were pre-treated with these agents and cultured in their absence. The agents did not stimulate colony development in the absence of colony-stimulating factor and enhancement of colony number occurred only in cultures containing a concentration of colony-stimulating factor which was sub-optimal in terms of maximum colony development. There was no indication of increased colony-stimulating factor production by treated marrow cells under the experimental conditions used to show colony enhancement. It was concluded that the agents caused an increased responsiveness of colony-forming cells to colony-stimulating factor.

Adenosine Monophosphate↗
PubMed1977

The in vitro differentiation of density sub-populations of colony-forming cells under the influence of different types of colony-stimulating factor.

The in vitro proliferation and differentiation of myeloid progenitor cells (CFU-c) in agar culture from CBA/Ca mouse bone marrow cells was studied. Density subpopulations of marrow cells were obtained by equilibrium centrifugation in continuous albumin density gradients. The formation of colonies of granulocytes and/or macrophages was studied under the influence of three types of colony-stimulating factor (CSF) from mouse lung conditioned medium CSFMLCM), post-endotoxin mouse serum (CSFES) and from human urine (CSFHu). The effect of the sulphydryl reagent mercaptoethanol on colony development was also examined. The density distribution of CFU-c was dependent on the type of CSF. Functional heterogeneity was found among CFU-c with partial discrimination between progenitor cells forming pure granulocytic colonies and those forming pure macrophage colonies. Mercaptoethanol increased colony incidence but had no apparent effect on colony morphology or the density distribution of CFU-c.

Animals↗
PubMed1978

Effect of myleran on murine hemopoiesis. III. Changes in the density distribution of spleen colony forming (CFU-S) and agar gel colony forming cells (CFU-C).

Spleen colony forming cells (CFU-S) and agar-gel colony forming cells (CFU-C) are separate but heterogeneous cell populations, as measured by buoyant density. Myleran (MY) abrogated the major (lighter density) components of the CFU-S and CFU-C compartments, thus shifting the surviving CFU-S and CFU-C density profiles into the higher density region. The normal spleen colony erythrocytic : granulocytic (E:G) ratio profile showed three density regions with different distributions of erythroid and granulocytic colonies. The preponderantly erythrocytic colony-generating CFU-S of the intermediate density regions were eradicated by MY. Comparison of the density distribution of erythrocytic and granulocytic colony-generating CFU-S of normal bone marrow showed that the erythrocytic CFU-S profile paralleled that of total CFU-S, while most of the granulocytic CFU-S were contained in the major (and lowest density) peak. MY eradicated the two main (and lowest density) peaks of CFU-S; surviving CFU-S occurred preponderantly in a minor (higher density) peak which has a high potential for generating erythrocytic colonies.

Animals↗
PubMed1976

Human granulocytopoietic colonies in diffusion chambers in mice: growth of colonies and the effect of host irradiation.

Normal human non-separated bone marrow cells were cultured in fibrin clots in diffusion chambers implanted intraperitoneally in mice, and harvested at different intervals by a previously described chamber centrifugation technique. This method demonstrates the presence of cell aggregates in the diffusion chambers. When the chambers are implanted in irradiated mice (450 R) and retransplantated into newly irradiated mice every seventh day, a continous increase in number of cells per granulocytopoietic aggregate is observed from day 8 to day 21. This is compatible with the view that the aggregate is observed from day 8 to day 21. This is compatible with the view that the aggregates are colonies. The term 'colony forming unit diffusion chamber' (CFUD) is suggested to denote the ancestor(s) of the colonies. However, formal proof that one colony is derived from one cell is lacking. Preirradiation of mice with 450 R significantly increases the number of neutrophilic granulocytopoietic colonies at day 14, provided the chambers are retransplantated to newly irradiated mice at day 7, indicating that the neutrophilic colony forming unit or its progeny is involved as at least one of the targets of the stimulating effect of host irradiation. In contrast, no effect of host irradiation on the numbers of eosinophilic colonies was observed.

Adult↗
PubMed1977

The statistical distribution of colony counts in the spleen colony assay.

The statistical distribution of colony counts in the spleen colony assay was determined in C57BL/6 and C3H mice under varying conditions of the assay procedure. Assay mice were given syngeneic bone marrow cells from normal or irradiated donors. The mice were injected intravenously with marrow cells either immediately or 3 days after irradiation. Eight days after engraftment, spleen colonies were counted and the statistical distribution of colony counts was determined using a mean to variance ratio criterion. Analysis of the data indicated that the statistical distribution of colony counts was different in the two strains of mice. In addition, delaying time of injection of mice with bone marrow cells or injecting bone marrow cells form irradiated donors also changed the statistical distribution. The significance of this work lies in the fact that modification of the usual assay procedure can alter the statistical distribution of colony counts, thus changing the statistical accuracy of the assay. Modification in the assay procedure, such that an increase occurs in the variance of the distribution of colony counts, must be compensated for by increasing the number of spleens counted in order to maintain a constant level of statistical accuracy in experiments.

Animals↗
PubMed1977

The effect of cyclic AMP on human colony forming cell and colony stimulating factor production.

The effect of cyclic AMP (cAMP) and related nucleotides on human colony forming cells (CFC) and those cells producing colony stimulating factor (CSF) was studied in vitro. When added at physiological concentrations (10(-2) to 1 micron), exogenous cAMP stimulated maximum colony formation in cultures without feeder layers. Related nucleotides stimulated colony formation to a lesser extent and in decreasing order of free energy. All nucleotides inhibited colony formation in concentrations above 1 micron. A velocity sedimentation cell separation technique was used to obtain cell fractions rich in CFC but poor in CSF-producing cells. Such fractions did not respond to cAMP stimulation. These studies suggest that exogenous cAMP stimulates human bone marrow to form colonies in vitro by increasing the release and/or production of endogenous CSF.

Adenine↗
PubMed1979

Bovine granulocyte/macrophage and erythroid colony culture: characteristics of the colonies and the assay systems.

Bovine bone marrow granulocyte/macrophage colonies were cultured in vitro in methyl cellulose and in plasma clots using bovine endotoxin-stimulated serum as a source of colony stimulating activity. The endotoxin-stimulated serum was four times as potent as the control serum in the methyl cellulose cultures. No significant increase in the number of colony forming units was observed when bovine marrow cells were maintained in suspension cultures for various periods prior to plating in methyl cellulose. The percentage of glass/plastic adherent cells in bovine marrow cells was observed to be 43% +/- 12 (SD). Benzidine positive erythroid colonies appeared in plasma clot cultures on day 4 and disappeared by day 9. No second population of erythroid colonies appeared either as a function of time or as a function of erythropoietin concentration. The optimum erythropoietin concentration for bovine erythroid cultures was found to be 1.0 unit/mL. A significant difference was observed between animals in their marrow capacity to produce erythroid colonies in culture but no significant difference was observed within individual animals over a period of three months.

Animals↗
PubMed1975

Haematopoietic defects of W/WV mice studied with the spleen colony, agar colony, and diffusion chamber techniques.

Bone marrow progenitor cells from anaemic W/WV mice were compared with normal +/+ cells utilizing the spleen colony, the agar colony and the diffusion chamber techniques. Spleen colony formation from W/WV cells was markedly defective, and more so for erythroid than for granuloid colonies. The progenitor cell concentration was apparently normal as measured by the two other techniques. The concentration of circulating progenitor cells also seemed to be normal. On the other hand, the cell formation per progenitor cell was subnormal in all three assay systems. The initial proliferative response of W/WV spleen colony-formers and agar colony-formers to short-term diffusion chamber culturing was apparently normal. The incorporation of 3H-thymidine, related to the number of proliferative granulocytes present in the chambers, also seemed to be normal. The results indicate that the W/WV defect is not limited to the multipotent stem cells. A possible interpretation is that it is the capacity for continued self-renewal of immature cells that is defective.

Anemia↗
PubMed1975

Stability of pathogenic colony types of Neisseria gonorrhoeae in liquid culture by using the parameters of colonial morphology and deoxyribonucleic acid transformation.

This investigation describes the surveillance of the colonial stability of the pathogenic type 1 from the gonococcal strain F62 to the nonvirulent types 3 and 4 in different liquid media. The maintenance of the colony types was monitored by the parameters of colonial morphology and deoxyribonucleic acid-mediated transformation. During growth in a complex medium, Mueller-Hinton broth, only 46.7% of the gonococcal population remained as type 1 after 12 h. The greatest change in the type 1 colony-forming units correlated with the decline in viable count. The conversion process could not be prevented by the continual maintenance of the gonococcus in logarithmic growth. The frequency of transformation from PRO(minus) (proline) to PRO(plus) was proportional to this decrease in type 1 colony-forming units. In contrast to Mueller-Hinton medium, the chemically defined minimal medium Gonococcal Genetic Medium (GGM) was capable of maintaining approximately 90% of the gonococcal population in the type 1 colonial form after 16 h of growth, despite a decrease in the viable count. Although the percentage of type 1 appeared to remain constant in GGM, the apparent transformation frequency increased approximately 24-fold from 0 to 12 h of growth. GGM appears to stimulate or maintain competence, as evidenced by an eightfold increase in transformation when cells are exposed to deoxyribonucleic acid in GGM as compared to Mueller-Hinton.

Culture Media↗
PubMed1978

The marrow colony forming cell and serum colony stimulating factor of the chicken.

Marrow cells of the chicken produced colonies in semisolid media. Developing colonies consisted of granulocytes, macrophages or a mixture of these two cell types. The granulocyte-macrophage CFC was nonadherent. An adherent 'CFC' was also present and it differed in several ways from the nonadherent CFC: (a) clones contained only macrophages, (b) they contained a core of nonrefractile cells, (c) their appearance was delayed 1-2 weeks, (d) they were unaffected by the presence of erythrocytes and (e) the efficiency of cloning was increased but the percentage of clones able to produce 50 or more cells was markedly decreased, i.e., the cluster/colony ratio was increased. The growth of both colony types was strictly dependent on the presence of CSF. Data obtained from dose-response studies on unfractionated marrow indicated that clusters and colonies were derived from single cells. The CSF of chicken serum yielded sigmoid dose-response curves when tested on marrow cells. Calf serum could not support cluster or colony formation when tested alone but it did have an enhancing effect on the CSF of chicken serum. Levels of serum CSF were increased by injecting chickens with bacterial endotoxin. This phenomenon occurred with five chicken lines tested, but certain chickens of the Kimber line did not respond to endotoxin with elevated levels of CSF.

Animals↗
PubMed1977

Bone marrow colony-forming cells in mice with virus-induced lymphoid leukemia: relation to serum colony-stimulating activity and blood granulocytes.

Mice with advanced lymphoid leukemia have elevated peripheral blood granulocytes and elevated serum colony-stimulating activity, which promotes the in vitro growth of granulocyte and/or macrophage colonies. The number of bone marrow precursor cells of the in vitro granulocyte and/or macrophage colonies varied from normal to 10% of normal. The elevation of colony-stimulating activity correlated best with a combination of increased blood granulocytes and a deficiency of bone marrow precursor cells, which suggested that colony-stimulating activity is a leukopoietin that increases the efficiency and rate of production of granulocytes.

Animals↗
PubMed1978

Studies on the morphology of colonies of bacilli of BCG-Poland substrain. I. The influence of iron on the type of BCG-Poland colonies.

The influence of iron concentration in Sauton's medium solidified with agar on the type of colonies of BCG-Poland substrains, BCG-Rio de Janeiro, BCG-France, BCG-Denmark and BCG-Japan substrains has been examined. Of all the studied BCG substrains only the BCG-Poland substrain formed rough (R) and smooth (S) colonies. In the investigated substrains rough colonies became smaller with the decrease of iron concentration but they retained their characteristic surface roughness. The smooth colonies which in the same conditions appeared only in BCG-Poland substrain did not display such dependency on iron concentration. When the incubation period was prolonged secondary rough colonies appeared among the smooth ones, regardless of the iron concentration in the medium.

BCG Vaccine↗
PubMed1979

Cyclic hematopoiesis. Effects of endotoxin on colony-forming cells and colony-stimulating activity in grey collie dogs.

Cyclic changes in blood neutrophil counts of grey collie dogs with cyclic hematopoiesis can be eliminated by daily endotoxin injections. Studies were performed to determine the mechanism whereby endotoxin alters this disease. Bone marrow granulocyte-macrophage progenitor cells (colony-forming cells [CFUc]) showed cyclic variation in the untreated grey collie, which was eliminated by chronic endotoxin treatment (Salmonella typhosa lipopolysaccharide W, 5 microgram/kg per day). Similar cyclic variation in blood CFUc was eliminated by this treatment. Tritiated thymidine suicide of the marrow colony-forming cells failed to show cyclic changes to explain the marked swing in CFUc numbers in untreated grey collies. The thymidine suicide rates were not significantly changed by chronic endotoxin treatment. Similarly, serum colony-stimulating activity did not show cyclic variation with the cyclic neutrophil counts in untreated grey collies and was not altered by chronic endotoxin treatment. We suggest that endotoxin eliminates neutrophil cycling in cyclic hematopoiesis by a direct effect on the flux of pluripotent stem cells into the committed stem cell compartment and that this occurs independent of changes in serum colony-stimulating activity.

Animals↗
PubMed1976

Relationship of colony-stimulating activity to apparent kill of human colony-forming cells by irradiation and hydroxyurea.

Suspensions of human bone marrow cells were subjected to 137Cs irradiation in vitro and then cultured in semisolid agar medium. Cultures of irradiated cells were stimulated with colony-stimulating activity (CSA) of different potencies, and it was found that the amount of stimulation applied to cultures influenced the apparent kill of colony-forming cells (CFC). It was also found that the effects of irradiation on colony formation were not confined to CFC kill since medium conditioned by cells during irradiation exhibited stimulatory and inhibitory properties after treatment by 600 and 1000 rads, respectively. Studies in which irradiated cells were pretreated with hydroxyurea indicated that CFC in the DNA synthetic phase of the cell cycle were particularly sensitive to low doses of irradiation. The proliferative capacity of CFC surviving 1000 rads was undiminished as judged by their ability to form large colonies. Estimates of CFC kill by hydroxyurea were also affected by the level of CSA.

Bone Marrow↗
PubMed1978

Specific antibody-forming B-lymphocyte colonies. I. Distribution and nature of SRBC antibody-forming B-lymphocyte colonies in mouse lyphomyeloid organs.

Wehn normal mouse spleen or lymph node cells are cultured for 7 days in agar-medium containing 2-mercaptoethanol and sheep red blood cells (SRBC), approximately one B lymphocyte colony (BLC) develops per 50-100 cells seeded. Incubation of cultures for 3 h with guinea-pig complement at day 7, demonstrated that 0.05-0.25% of all BLC form specific antibody against SRBC (SRBC-AF-BLC). The SRBC specific colonies appear centrally in lytic plaques of 2-5 mm in diameter and cells recovered from individual SRBC-AF-BLC were shown to produce antibodies of the IgM class against SRBC. Cells forming SRBC-AF-BLC are absent in the new-born and infantile spleen but appear in adult mice with a frequency of 5, 10-25 and 25-70 per 10(6) bone marrow, spleen and lymph node cells respectively. specific immunization in vivo or in vitro does not greatly affect the number of SRBC-AF-BLC-forming cells. Cytolysis of spleen cells with anti-Ig serum plus complement prior to culture reduced the number of total BLC and that of specific SRBC-AF-BLC by 93% and 94% respectively. The peak sedimentation velocity of both SRBC-AF-BLC-forming cells and total BLC-forming cells was 3.5 mm/h. Spleen cells enriched 200-300 times for cells that bind specifically to the hapten NIP were not enriched for cells forming colonies with specific antibody production against NIP. The data indicate that the cells that give rise to specific antibody-forming colonies belong to a mature virgin B cell group of small Ig-positive B lymphocytes.

Age Factors↗