Separation of rabbit marrow precursor cells by combined isopycnic
sedimentation and electronic cell sorting
RB Scott, WM Grogan and JM Collins
Separation of developing cells into fractions of differing stages of
maturity is critical to effective biochemical study of the process of
cellular differentiation. Density gradient techniques utilizing rate- zonal
or isopycnic separations have permitted partial separations based on cell
mass or cell density. In this study the separation of various rabbit marrow
cells was improved by high-speed flow analysis and sorting in a Coulter
Two-Parameter Cell Sorter. After preliminary isopycnic separation of marrow
cells in Ficoll-Hypaque, cells were sorted into lymphoid and myeloid
elements, utilizing light-scatter (LS) profiles to determine sorting.
Characteristic LS patterns were present for erythrocytes, lymphocytes,
devitalized cells, and granulocytes. When erythrocytes and their precursors
were removed by hypotonic lysis, the remaining granulocytes could be sorted
to give samples with much greater purity with respect to developmental
stage than was possible with density gradients alone. Thus the combination
of techniques represents a significant improvement in the ability to study
the mechanisms of normal or altered cellular maturation.
Volume 51,
Issue 6,
pp. 1137-1148,
06/01/1978
Copyright © 1978 by The American Society of Hematology
