Preparation of Platelet Free Mononuclear Cells

Abstract

Platelets are similar in density to PBMC and thus are contaminants of PBMC isolated by Ficoll density gradient centrifugation the established procedure for preparing PBMC. Platelets are very sticky and often interfere with experiments requiring lymphocytes and or monocytes (PBMC). Methods, compositions and kits are provided for the rapid removal of the undesired platelets from PBMC yielding platelet free PBMC in high yield. The selection process, in the preferred embodiment of the invention, is based on magnetic metallic particles that are uniquely suited for removal of platelets from whole blood or PBMC. The preferred particle is composed of nickel metal in a preferred size range of 0.8-3.5 micron. Specific anti-platelet binding agents are bound to the nickel particles. The particles are mixed with the sample rapidly without non-specifically binding to non-targeted cells. The sample is then placed in a magnetic field and the platelets bound to the magnetic particles are very rapidly removed yielding platelet free whole blood or PBMC in very high yield for further biological experimentation. The method also provides for the removal of platelet and platelet fragments from PBMC preparations where platelets may be present prior to freezing or after thawing the PBMC preparation using a three-bead combination anti-platelet magnetic particle.

Dependencies are inferred from the claim text.