Homogeneous monocytes and macrophages from hES cells following coculture-free differentiation in M-CSF and IL-3.

In the present study we aimed at producing homogeneous populations of monocytes and macrophages from human embryonic stem (hES) cells. Human embryonic stem cell lines KCL001, KCL002, and HUES-2 were differentiated into monocytes by coculture-free differentiation with two growth factors using a three-step method. The method involved embryoid body (EB) formation in hES media, directed differentiation with macrophage colony-stimulating factor and interleukin (IL)-3, and harvest of nonadherent monocytes from the culture supernatants. Transcriptome analysis of the esMC and the esMDM revealed a distinct myeloid signature that correlated with primary adult blood-derived monocytes and spleen tissue samples but not with other tissue samples tested. In conclusion we have developed a simple and efficient method for producing homogeneous populations of monocytes and macrophages from hES cells. esMCs have a myeloid signature and can differentiate into functional macrophages. The method should prove useful in answering experimental questions regarding monocyte and macrophage development and biology. Triplicates of monocytes and macrophages derived from human embryonic stem cells were compared to a triplicate of freshly isolated human blood monocytes.