Figure 1

Derivation of macrophages from lentiviral vector transduced and normal hES cells. A) Transduced and non-transduced H1 hES cells were cultured on mouse S17 bone marrow stromal cell layers to derive cystic bodies. Cystic body derived CD34 cells were purified by positive selection with antibody conjugated magnetic beads and placed in methocult media to obtain myelomonocytic colonies. Pooled colonies were cultured in liquid cytokine media supplemented with GM-CSF and M-CSF to promote macrophage growth. For comparison, fetal liver derived CD34 cells were cultured similarly to derive macrophages. Representative ES cell colonies, cystic bodies, methocult colonies, and derivative macrophages are shown with GFP expressing cells fluorescing green under UV illumination. B) Representative FACS profile of hES cell derived CD34 cells stained with PE conjugated antibodies. Percent positive CD34 cells are shown with isotype control shown in the left panel.