Development of a lacZ marked WEHI-3B D+ murine leukemic cell line as an in-vivo model of acute non-lymphocytic leukemia.
Established leukemic cell lines have been useful models for studying the biology of leukemia. Analysis of the actions of differentiating agents on leukemic cell lines in vivo has been limited by an inability to unambiguously distinguish host hematopoietic elements from differentiated leukemic cells. In order to identify and quantify leukemic cells during in vivo studies, a derivative of the murine myelomonocytic leukemia cell line WEHI-3B D+, which stably expresses beta-galactosidase, was constructed utilizing retroviral vector gene transfer. This cell line, termed WEHI-3B D+/lacZ 2.8, demonstrated in vitro growth and differentiation properties similar to the parental cell line. WEHI-3B D+/lacZ 2.8 expressed high levels of beta-galactosidase following prolonged in vitro growth and following differentiation in suspension cultures and clonogenic assays. In vivo, WEHI-3B D+/lacZ 2.8 was leukemogenic and high level expression of beta-galactosidase was maintained. Quantification of tissue involvement with WEHI-3B D+/lacZ 2.8 leukemia was performed utilizing staining with the fluorogenic beta-galactosidase substrate fluorescein di-beta-galactoside and fluorescence-activated cell sorting analysis. In vivo differentiation efficiency following granulocyte colony-stimulating factor (G-CSF) administration was determined using a simultaneous nuclear and cytoplasmic staining procedure. Results indicate that treatment of mice inoculated with WEHI-3B D+/lacZ 2.8 cells with G-CSF administration causes detectable but limited differentiation.
Smith
C
C
Division of Hematologic Oncology, Memorial Sloan Kettering Cancer Center, New York, NY.
Muench
M O
MO
Knizewski
M
M
Gilboa
E
E
Moore
M A
MA
D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics; D002455:Cell Division; D004926:Escherichia coli / Q000235:genetics*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000627:therapeutic use; D015479:Leukemia, Myelomonocytic, Acute / Q000201:enzymology* / Q000401:mortality / Q000473:pathology / Q000628:therapy; D017254:Leukemic Infiltration; D051379:Mice; D008807:Mice, Inbred BALB C; D012190:Retroviridae / Q000235:genetics*; D012680:Sensitivity and Specificity; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay; D001616:beta-Galactosidase / Q000032:analysis* / Q000235:genetics
D014408:Biomarkers, Tumor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D001616:beta-Galactosidase