SERUM-FREE MEDIUM FOR THE CULTURE OF HUMAN HEMATOPOIETIC CELLS

THE OBJECTIVES OF THIS PROJECT ARE TO DEVELOP A STANDARD SERUM-FREE MEDIUM THAT WILL SUPPORT THE PROLIFERATION AND DIFFERENTIATION OF HUMAN BONE MARROW STEM CELLS IN BOTH SHORT-TERM AND LONG-TERM CULTURES. THE RESEARCH WILL FOCUS ON THE DEVELOPMENT OF A SERUM-FREE FORMULATION TO SUPPORT THIS PROLIFERATION AND DIFFERENTIATION IN SHORT-TERM CULTURES OF GRANULOCYTES, MONOCYTES, EOSINOPHILS, ERYTHROID PRECURSORS, CFU-E, AND BFU-E. THE INITIAL SERUM-FREE FORMULATION WILL BE ONE THAT SUPPORTSTHE GROWTH OF HUMAN GRANULOCYTE AND MONOCYTE COLONIES. THE LONG-TERM BONE MARROW STUDIES WILL FOCUS ON THE DEVELOPMENT OF A SERUM-FREE FORMULATION THAT WILL SUPPORT THE PROLIFERATION OF THE STROMAL CELLS. THE INITIAL SERUM-FREE MEDIUM WILL BE FORMULATED FOR MURINE LONG-TERM BONE MARROW CULTURES. ONCE THIS IS ACHIEVED, THE STROMAL LAYER, GROWN IN SERUM-FREE MEDIUM, WILL BE ANALYZED FOR ITS ABILITY TO SUPPORT THE PROLIFERATION AND DIFFERENTIATION OF MYELOID STEM CELLS. SPECIFIC ATTENTION WILL BE PAID TO THE PURITY OF EACH COMPONENT AND THE USE OF ONLY HUMAN COMPONENTS WHERE BIOLOGICAL COMPONENTS ARE NECESSARY. THESE STUDIES WILL EMPLOY IN VITRO ASSAYS THAT ARE ROUTINELY USED FOR HEMATOPOIETIC CELLS.