Detection of in vivo differentiation of murine WEHI-3B D+ leukemia cells transfected with the lac-Z marker gene using two-color flow cytometry.
The in vivo induction of the differentiation of murine WEHI-3B D+ myelomonocytic leukemia cells was measured by flow cytometry, simultaneously staining leukemia cells for the marker exogenous beta-galactosidase and for differentiation by the antigen Mac-1 (CD11b/CD18). The WEHI-3B D+ leukemia cells were transfected with the E. coli lac-Z gene by electroporation and subclones that constitutively expressed high levels of the lac-Z gene product beta-galactosidase were established. Flow cytometric analyses of cells in the peritoneal cavities of mice bearing leukemia cells showed that cells continued to express beta-galactosidase for at least 14 days, and they were distinguishable from host-derived cells in vivo by their expression of the transfected gene. Simultaneous determination of the beta-galactosidase activity and Mac-1 content of cells in the peritoneal cavities of mice revealed that administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) to the mice enhanced the expression of Mac-1 antigen by beta-galactosidase-positive cells. The results demonstrate that G-CSF may have clinical potential as a therapeutic differentiating agent, and that flow cytometric analysis provides a useful in vivo system to evaluate the therapeutic potential of agents capable of inducing terminal differentiation.
Hayashi
M
M
Department of Chemotherapy, Saitama Cancer Center Research Institute, Japan.
Tomida
M
M
Hozumi
M
M
King
I C
IC
Sartorelli
A C
AC
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D005434:Flow Cytometry / Q000379:methods; D005819:Genetic Markers; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D007763:Lac Operon; D015479:Leukemia, Myelomonocytic, Acute / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D014162:Transfection
D005819:Genetic Markers; D016179:Granulocyte Colony-Stimulating Factor