Flow cytometric analysis of megakaryocytopoiesis: strategy for defining mitotic and endoreduplicative events in the committed progenitor compartment.
A new cellular approach to the analysis of the physiology and regulation of the megakaryocyte progenitor cell compartment is proposed. It utilizes initial progenitor cell separation by centrifugal elutriation followed by flow cytometry and viable cell sorting using quantitative, vital DNA stains and secondary cytoplasmic and/or membrane markers to isolate megakaryocyte progenitors on the basis of DNA content for subsequent analysis of proliferative (mitotic) activity in an in vitro clonogenic assay. It should be possible to delineate whether mitosis (as measured by the number of CFU-M per culture) is limited to diploid cells and determine the distribution of CFU-M in subclasses of cells identified by the secondary label. This kind of study will provide new information on the relationship between progenitor cell size, DNA content and proliferative activity within the framework of a working model of progenitor cell kinetics.
D000818:Animals; D002469:Cell Separation; D003584:Cytological Techniques; D004247:DNA / Q000032:analysis; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008533:Megakaryocytes / Q000166:cytology*; D008938:Mitosis; D008954:Models, Biological; D012377:Rodentia
D004247:DNA