The objectives of this proposal will be 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 culture. The research will focus on the development of a serum-free formulation(s) that will support the proliferation and differentiation in short-term culture of granulocytes, monocytes, eosinophils, erythroid precursors, CFU-E and BFU- E. The initial supports the growth of human granulocyte and monocyte colonies. The long-term bone marrow studies will focus on the development of a serum-free formulation(s) that will support the proliferation of the stromal cells. The initial serum-free medium utilized will be one that we have 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 are necessary. Such studies will employ in-vitro assays that are routinely used for hematopoietic cells. Such a medium will be useful in standardizing in-vitro hematopoietic assays and in-vitro expansion of specific populations of bone marrow stem cells.
