Identification of murine haemopoietic stem cells with monoclonal antibodies

textabstractThe work described in this thesis deals with the production and application of monoclonal antibodies to characterize murine haemopoietic stem cells. Monoclonal antibodies directed against determinants uniquely expressed on various classes of haemopoietic stem cells should provide valuable tools for analyzing the early differentiation steps in the stem cell compartment at the molecular level, for elucidating the relationhip between the various stem cell classes and for purifying the various stem cell types. In the following chapter the experimental approaches used are described. Chapter 3 deals with attempts to produce stem cell specific monoclonal antibodies. The characterization of the monoclonal antibodies and their possible usefulness in haemopoietic studies are described in chapter 4. Using one of the produced monoclonal antibodies a widely applicable multiparameter flow cytometric procedure for the identification and purification of murine haemopoietic stem cells is described in chapter 5. In chapter 6 various purification procedures for haemopoietic stem cells are compared. It appeared that the highly purified day-12 CFU-S suspensions are heterogeneous, therefore the usefulness of the CFU-S assay as endpoint-assay for pluripotent haemopoietic stem cells is discussed. In the final chapter, the results will be considered in a broader perspective and possible approaches for further study will be outlined