A new method for separating mononuclear cells from whole blood.

Abstract

The LeucoPREP tube, a new method for separating mononuclear cells from whole blood, is described. The LeucoPREP tube is a 7-ml glass tube which contains a polyester gel and a solution of a sucrose polymer and sodium diatrizoate (Histopaque, Sigma). Platelets, lymphocytes, and monocytes are separated from granulocytes and erythrocytes based on differences in density. Platelets are then removed from the mononuclear cells by washing. The yield, viability, and percentage of various cell subpopulations obtained from the LeucoPREP tube are equivalent to those obtained from standard Ficoll-based methods. However, the LeucoPREP tube is a faster, less technically demanding method than Ficoll. The major advantages of the LeucoPREP tube are that 1) whole blood is added directly into the tube, unlike Ficoll methods, in which the blood must be diluted and carefully layered; and 2) it requires a very short (10 min) centrifugation time. The LeucoPREP tube should be highly desirable to anyone processing many blood samples, for example, clinical laboratories.