CD34⁺CD90⁺ cells exposed to 5azaD and TSA are capable of differentiating into multiple blood cell lineages. Displayed is the schema used to evaluate the differentiation potential of CD34⁺CD90⁺ cells after exposure to 5azaD and TSA. After 9 days of initial culture (described in “Materials and methods”) cells were sorted into 2 subsets, CD34⁺CD90⁺ and CD34⁺CD90^-, and then placed in a secondary culture supplemented with IL-3, IL-6, GM-CSF, TPO, SCF, and EPO. The phenotype of cells was determined by staining with monoclonal antibodies directed toward CD41, CD71, CD11b, CD14, CD34, and CD90. Each panel shows a representative flow cytometric profile with each of these monoclonal antibodies: (A) isotype matched control; (B) CD11b and CD14; and (C) CD34⁺ and CD90⁺. (D) mAb stain with CD71 (dotted line is the matched isotype control; the continuous line represents CD71 mAb staining). (E) Single antibody stain with CD41 (dotted line is the matched isotype control; the continuous line represents CD41 mAb staining). (F-G) Cytospin preparations were stained with Giemsa and Wright stain and were viewed with a light microscope (original magnification × 400). Each panel is represented as follows: (F) CD34⁺CD90⁺ cells after 16 days of culture with pretreatment with 5azaD and TSA; and (G) marrow CD34⁺ cells cultured for total of 16 days, not exposed to 5azaD and TSA.