Density gradient segregation of bone marrow cells with the capacity to form granulocytic and macrophage colonies in vitro*1

Janoshwitz, H.; Moore, M. A. S.; Metcalf, Donald

doi:10.1016/0014-4827(71)90608-2

Cited by 24 publications

(4 citation statements)

References 8 publications

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“…Thus, heterogeneity of the day 7 M-CFC exists both in the expression of the YBMi42 antigen and in their response to different growth factors. A similar heterogeneity in response to growth factors has been observed previously with bone marrow cells using sedimentation velocity techniques (Janoshwitz et al, 1971;Byrne et al, 1977;Bol et al, 1979).…”

Section: Discussionsupporting

confidence: 80%

Segregation of mouse hemopoietic progenitor cells using the monoclonal antibody, YBM/42

Watt

Gilmore

Metcalf

et al. 1983

Journal Cellular Physiology

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A rat monoclonal antibody, YBMi42, directed against mouse leukocyte common antigen, was used for the analysis and separation of hemopoietic progenitor cells from mouse bone marrow and fetal liver. Cells were fractionated o n a FACS-II cell sorter and the resulting subpopulations examined for their morphology and ability to form colonies in agar (for day 7 colonies) and methylcellulose (for day 2 erythroid clones). The antibody bound to all leukocytes, including blast cells and day 7 hemopoietic progenitor cells (day 7 colony forming cells, CFC), but not to erythrocytes or nucleated erythroid cells. This antibody can be used to advantage to enrich for early progenitor cells from mouse fetal liver, in which the majorityof cells (70%) are nucleated erythroid cells. In day 12 fetal liver, approximately 10% of all cells bind this antibody strongly and, of these approximately 70% are blast cells. Contained within this positive population are 95% of all day 7 CFC. In the most enriched fraction about 20% of t h e cells formed day 7 colonies. This represents a 25-fold enrichment over unsorted fetal liver. The negative fractions contain 94% of all cells forming erythroid clones ( 2 8 cells) o n day 2 of culture (day 2 CFU-E). In the most enriched fraction, 20% of the cells are day 2 CFU-E. Day 7 CFC can therefore be well separated from day 2 CFU-E, with good recovery of both cell types, by use of a single label. Day 7 colony forming cells were classified as granulocyte (G-CFC), macrophage (M-CFC), mixed granulocyteimacrophage (GM-CFC), pure erythroid (El, or mixed erythroid (E,J. A high enrichment for multipotential cells is achieved and constitutes 3-5% of cells in the most enriched fraction. Most types of day 7 CFC could not be separated with YMBi42, but GM-CFC and M-CFC exhibit a broader distribution than t h e other CFC with regard to fluorescence intensity. This implicit heterogeneity in GM-CFC and M-CFC is further substantiated by the finding that myeloid progenitors in the different FACS fractions also share a differential reactivity to different sources of growth factors.

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Section: Discussionsupporting

confidence: 80%

Segregation of mouse hemopoietic progenitor cells using the monoclonal antibody, YBM/42

Watt

Gilmore

Metcalf

et al. 1983

Journal Cellular Physiology

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“…Colonies are initially granulocytic but with longer incubation can transform to macrophages (Metcalf,197Ib). Density separation has demonstrated heterogeneity among CFU-c with respect to the type of colony they form under the same culture conditions (Janoshwitz et al, 1971). This was seen here with both types of CSF.…”

Section: Discussionsupporting

confidence: 67%

Stimulation of in Vitro Granulocyte‐Macrophage Colony Formation by Mouse Heart Conditioned Medium

Byrne¹,

Heit²,

Kubanek³

1978

Br J Haematol

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Medium conditioned by mouse heart tissue was found to contain colony-stimulating factor(s) (CSF) which produced in vitro colonies of granulocytes and/or macrophages with bone marrow cells of CBA mice. The action of this CSF type (CSFMHCM) was compared with that of post-endotoxin mouse serum (CSFES on colony formation by density subpopulations of colony-forming cells (CFU-c), separated by isopycnic centrifugation in a continous albumin density gradient. The number, size and morphology of colonies were examined. Mouse heart conditioned medium produced slightly more colonies and of larger size than endotoxin serum but the density distribution of their CFU-c were similar. Low density CFU-c produced predominately pure granulocytic colonies and high density ones predominately pure macrophage colonies with both types of CSF. There was no apparent relationship between colony size and CFU-c density.

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“…One might speculate that initially conditions in the DC were such as to favor a maximum differentiation of CFC into the granulocyte line, emphasizing the relationship of CFC to granulocyte formation rather than to macrophage formation. Using a density separation procedure it has been demonstrated that some CFC are intrinsically more likely to differentiate into granulocytic than into mixed or macrophage colonies (Janoshwitz, Moore & Metcalf, 1971). The data available on the morphology of the h ritro colonies derived from F3 and control cells, however, suggests that this type of selective separation of CFC was not achieved in our experiments.…”

Section: Discussionmentioning

confidence: 64%

Growth of Stem Cell Concentrates in Diffusion Chambers (Dc)

et al. 1974

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The growth in diffusion chamber (DC) of normal murine marrow and marrow separated by discontinuous albumin centrifugation was studied. The colony‐forming cells (CFC) assayed in soft agar, total cell counts and differentials were measured in the DC over a 19 day period after intraperitoneal implantation into CF1 mice. Growth of implants of normal marrow or fraction 3 (F3) in which CFC had been concentrated 1.7–3.9‐fold were compared at an initial cell concentration of 1 × 105. There was a good correlation between the number of CFC implanted with granulocyte production but not with macrophage production. When higher cell concentrations of normal unfractionated marrow were implanted growth was reduced as was recovery of CFC. In two experiments in which both CFU and CFC concentrations were measured there was a general correlation between the two.

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Density gradient segregation of bone marrow cells with the capacity to form granulocytic and macrophage colonies in vitro*1

Cited by 24 publications

References 8 publications

Segregation of mouse hemopoietic progenitor cells using the monoclonal antibody, YBM/42

Segregation of mouse hemopoietic progenitor cells using the monoclonal antibody, YBM/42

Stimulation of in Vitro Granulocyte‐Macrophage Colony Formation by Mouse Heart Conditioned Medium

Growth of Stem Cell Concentrates in Diffusion Chambers (Dc)

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