Flow cytometry data and methylcellulose culture. (A) Flow cytometry data from a secondary sheep, at 13 months after transplantation, that received a transplant of 1 × 10⁶ bone marrow cells harvested from a primary sheep that received a transplant of 100 000 CD34⁺/Lin^- cells derived from hESCs. Samples were collected before and 4 days after a GM-CSF treatment at 5 μg/kg/d for 5 days. (B) Methylcellulose culture of bone marrow cells from a secondary sheep that received a transplant of 1 × 10⁶ bone marrow cells harvested from a primary sheep that received a transplant of 100 000 CD34⁺/Lin^- cells derived from hESCs. Cultures were grown in media including GM-CSF and erythropoietin, and (i-ii) 2 typical granulocyte macrophage colony-forming unit (CFU-GM) colonies are depicted on day 14 (original magnification, × 40). Images were acquired using an Olympus IX71 microscope (Olympus, Melville, NY) with an Olympus Plan 4 × 0.01 numeric aperture objective lens, an Olympus DP70 camera, and Olympus DP Controller 2.1.1.183 software. Images were processed using Adobe Photoshop CS (Adobe Systems, San Jose, CA). (iii) Colonies were tested for human DNA (PCR for human beta-2–microglobulin) before and 3 days after a 5-day GM-CSF stimulation at 5 μg/kg/d. Three of 24 and 12 of 40 colonies tested positive for human DNA before and after the GM-CSF treatment, respectively. (Top) Human beta-2–microglobulin. (Bottom) Beta actin common to sheep and humans. Human DNA is shown in the last lane and methylcellulose colonies in lanes 1 to 8.