Separation of hemopoietic cells from adult mouse marrow by use of
monoclonal antibodies
T Hoang, D Gilmore, D Metcalf, S Cobbold, S Watt, M Clark, M Furth and H Waldmann
Primitive hemopoietic progenitor cells from adult mouse marrow have been
substantially enriched by virtue of a negative selection procedure with
monoclonal antibodies. It has been possible to segregate erythroid
progenitor cells at distinct stages of differentiation on the basis of
their cell surface antigens. This has been achieved with two monoclonal
antibodies reactive with the mature elements of bone marrow. YBM 34.3 binds
to a heat-stable antigen expressed on B lymphocytes, neutrophils, and cells
of the erythroid lineage. YBM 6.1 reacts with cells of the neutrophil,
eosinophil, and monocyte series but does not bind to colony- forming cells.
Separation is achieved by indirect immunoadsorption (panning) with YBM 34.3
on Protein-A-coated plastic plates followed by FACS II cell sorting with
YBM 6.1. The combined procedures yield a marrow population containing 58%
immature cells (blasts, promyelocytes, and myelocytes) and 9.5% clonogenic
cells. In addition, differential binding of YBM 34.3 can be used to
segregate erythroid progenitor cells at distinct stages of differentiation
(day 7 BFU-E, day 5 BFU-E and CFU- E) either by cell sorting or panning. It
is shown that both techniques give a comparable degree of resolution of the
different cell types with, however, an appreciable advantage of panning
over cell sorting in allowing the rapid handling of large numbers of cells.
Volume 61,
Issue 3,
pp. 580-588,
03/01/1983
Copyright © 1983 by The American Society of Hematology
