Stem cells are the agents of hematopoietic reconstitution in bone marrow transplantation and are a target for gene therapy. This proposal is to facilitate human stem cell research by providing a rapid, high yield system for isolating and recovering stem cells from bone marrow, cord blood or peripheral blood sources. This system consists of a two-step process of specific sequential depletions of non-stem cell components using an erythrocyte precipitation buffer and an immunoaffinity column. This system is capable of processing large numbers of cells to produce high levels of enrichment for CD34+ cells. Preliminary results with both bone marrow and cord blood using this method have demonstrated a >70-fold increase in stem cell concentration and >70% recovery of total CFU activity. The initial focus of the study will be to optimize the procedure for enrichment of stem cells from different sources and determine the recovery of colony forming cells. This study will lead to the development of a product intended for the basic and clinical research market and will also form the basis for a phase II submission intended towards development of this methodology as a closed system medical devise.National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
