Demonstration of permanent factor-dependent multipotential (erythroid/neutrophil/basophil) hematopoietic progenitor cell lines.

Abstract

Multipotential hematopoietic progenitor cell lines have been established from nonadherent cell populations removed from continuous mouse bone marrow cultures. Clonal sublines of lines B6SUtA or B6JUt derived from single cells formed mixed colonies containing erythroid cells, neutrophil-granulocytes, and basophil/mast cells in semisolid medium containing erythropoietin and conditioned medium from pokeweed mitogen-stimulated spleen cells. Each of several subclones of cell line Ro cl formed colonies containing eosinophils, neutrophil-granulocytes, and basophil/mast cells in semisolid medium. Multipotentiality was maintained in vitro for over 2 1/2 years. In contrast, cell line 32D formed basophil/mast cell colonies with no detectable differentiation to other pathways. Multipotential cell lines did not produce detectable spleen colonies (CFUs) in vivo, nor did intravenous inoculation of up to 5 X 10(7) cells protect lethally irradiated mice from bone marrow failure. Newborn and adult mice inoculated with 5 X 10(7) cells showed no detectable leukemia or solid tumors after one year. Both multipotential and committed basophil/mast cell lines demonstrated absolute dependence upon a source of a growth factor(s) found in medium conditioned by WEHI-3 cells. These cell lines should be of value in studies of the regulation of hematopoietic stem cell differentiation in vitro.

The EVI1 gene in myeloid leukemia

“…32Dc13 cells are myeloid precursor cells which differentiate into granulocytes in response to G-CSF. 29 Although these cells normally express the endogenous Evi1 due to retroviral integration, 30 they will differentiate into granulocytes in response to G-CSF. 29 However, infection or transfection of 32Dc13 cells with constructs expressing human or murine EVI1 cDNA leads to block of G-CSF-dependent differentiation, and to inability of the cells to express myeloperoxidase and to differentiate to granulocytes.…”

“…29 Although these cells normally express the endogenous Evi1 due to retroviral integration, 30 they will differentiate into granulocytes in response to G-CSF. 29 However, infection or transfection of 32Dc13 cells with constructs expressing human or murine EVI1 cDNA leads to block of G-CSF-dependent differentiation, and to inability of the cells to express myeloperoxidase and to differentiate to granulocytes. 27,28 Expression of additional EVI1 in erythroid progenitors can also result in the loss of erythropoietin responsiveness.…”

Granulocytic differentiation of myeloid progenitor cells by p130, the retinoblastoma tumor suppressor homologue

“…Expression of the pRB family of pocket proteins during granulocytic di erentiation 32Dcl3 cell is a myeloid cell line derived from normal mouse bone marrow cells and is dependent on interleukin 3 (IL-3) for growth and survival (Greenberger et al, 1983). As previously reported, 32Dcl3 cells cultured in medium containing G-CSF in the absence of IL-3 cease proliferation and undergo granulocytic di erentiation, which is morphologically characterized by nuclear segmentation (Valtieri et al, 1987) (Figure 1a,b).…”

“…32Dcl3 is a non-tumorigenic myeloblastoid cell line established from mouse bone marrow and is totally dependent on interleukin-3 (IL-3) for its proliferation and survival (Greenberger et al, 1983). When the 32Dcl3 cells are treated with G-CSF in the absence of IL-3, they exit from the cell cycle and terminally di erentiate to granulocytes.…”

Vaccination with leukemia cells expressing cell-surface-associated GM-CSF blocks leukemia induction in immunocompetent mice

“…BCR-ABL þ 32D cells were obtained from the laboratory of Bruno Calabreta. The 32D cell line (clone 3) was derived from C3H/HEJ mice (Greenberger et al, 1983), and human BCR-ABL oncogene (b3/a2)gene was transduced into 32D cell line (Daley et al, 1990). BCR-ABL þ 32D cells induce leukemia on injecting 6-8-week-old C3H/HEJ mice (Matulonis et al, 1995).…”