Indirect differentiation of mouse embryonic stem cells into macrophage-like cells

Abstract

Macrophages derived from pluripotent stem cells represent an attractive cellular system for the study of lentiviruses. The possibility of obtaining cells with a phenotype similar to macrophages in the process of differentiation of mouse embryonic stem cells (ESCs) in the presence of the conditioned medium (CS) from bone marrow-derived stromal cells or interleukin 3 (Il-3) and macrophage colony-stimulating factor (M-CSF) was demonstrated. The results of flow cytofluorimetry showed that the replacement of CS with recombinant cytokines in induction media contributed to an increase in the number of cells positively stained antibodies (AB) against AG CD34 by 8% and CD45 by 20%. When seeding such cells in a methylcellulose medium supplemented with CS or a mixture of Il-3 and M-CSF, the formation of colonies with different morphology, including monocyte/macrophage, was observed with an efficiency of 0.11 % and 0.33%, respectively. The percentage of positively stained cells AB against F4/80 AG was 8±0.02 %. The cells demonstrated the ability to non-specific phagocytosis of latex particles. Thus, it is shown that it is possible to obtain cells with a phenotype similar to macrophages from mouse ESCs during differentiation, and the advantage of using two recombinant cytokines for this purpose is established.