The 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