Canine megakaryocytopoiesis: analysis utilizing a monoclonal antibody to a 140-kd dog platelet protein.
The dog, a convenient and relatively large animal whose size permits repetitive blood and marrow sampling, marrow biopsy, and mechanical apheresis, would be a suitable experimental model for the study of in vivo megakaryocytopoiesis. A monoclonal antibody to a 140-kd dog platelet membrane protein has been developed that reacts with canine megakaryocytes. Using the fluoresceinated derivative of this antibody to identify megakaryocytes and propidium iodide staining to measure relative DNA content, the DNA distribution of megakaryocytes in dog bone marrow or in cultured dog marrow cells could be rapidly assessed by flow cytometry. Normal dogs showed a modal ploidy of 16N (54%), with 17% 8N and 16% 32N. In contrast, dogs made thrombocytopenic by plateletpheresis showed a shift in distribution to higher ploidy cells (36% 32N). The data show that use of a specific marker of megakaryocytes in combination with flow cytometric analysis is an accurate and reproducible method of assessing megakaryocytopoiesis in a convenient and easily manipulable animal model.
Burstein
S A
SA
Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City 73190.
Friese
P
P
Downs
T
T
Epstein
R E
RE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000276:immunology / Q000378:metabolism*; D001798:Blood Proteins / Q000276:immunology*; D002455:Cell Division; D003584:Cytological Techniques; D004285:Dogs; D005434:Flow Cytometry; D008533:Megakaryocytes / Q000166:cytology*; D008970:Molecular Weight; D010983:Plateletpheresis; D012016:Reference Values; D013997:Time Factors
D000911:Antibodies, Monoclonal; D001798:Blood Proteins