A MONOCLONAL-ANTIBODY TO PLATELET GLYCOPROTEINS IIB AND IIIA COMPLEX - ITS USE IN PURIFYING HUMAN MEGAKARYOCYTES FROM STERNAL BONE-MARROW ASPIRATES FOR IMMUNOFLUORESCENCE STUDIES OF IA-LIKE ANTIGENS

  • 1 January 1983
    • journal article
    • research article
    • Vol. 11  (3) , 169-177
Abstract
A monoclonal antibody (PBM 6.4) to platelet and megakaryocyte glycoproteins IIb and IIIa was obtained and used to purify human megakaryocytes from sternal bone marrow aspirates by a simple method, consisting of a Percoll gradient centrifugation followed by affinity adherence on PBM 6.4-coated plastic surfaces, panning. Megakaryocytes, 80-90% pure and morphologically well preserved, were attached to poly-L-ornithine-coated multi-well microscope slides and immunofluorescence was done using monoclonal antibodies to human Ia-like antigens (DR, DC1). A higher proportion of DR-positive megakaryocytes was found, in comparison to the values reported by others, while DC1 antigen was detected on .apprx. 20% of megakaryocytes. The method described has the unique feature of enriching cells of the human megakaryocytic lineage from simple diagnostic sternal aspirates in an amount adequate for immunofluorescent and morphological analysis.