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UNI-SEPVAC # U-07

FOR DENSITY GRADIENT SEPARATION OF LYMPHOCYTES

 

INTRODUCTION

Density gradient centrifugation is the method of choice for isolation of lymphocytes. The success of the procedure, i.e. the recovery of viable lymphocytes with the lowest proportion of contaminating granulocytes and erythrocytes, depends to a large extent on the careful layering of the blood sample onto the separation medium and the maintenance of a sharp interface between the two solutions prior to centrifugation. To this end UNI-SEP* was developed.


The UNI-SEPVAC system allows the blood sample to be drawn directly into the centrifuge tube with no special precautions required. Thus, a large number of samples may be handled at the same time with no manipulation of the blood.

 


PRINCIPLE

UNI-SEPVAC is a sterile plastic centrifuge tube containing 3 ml of NycoprepTM (density 1.077 g/ml, osmolality 295 mOsm), which is sequestered in the bottom of the tube by a one-way plastic insert. Air has been removed from the tube to allow automatic filling with 4 ml of blood. Lithium heparin or sodium citrate is included in the upper chamber as anticoagulant. Blood drawn onto the insert does not disrupt the surface of the NycoprepTM layer. During centrifugation the insert opens to permit mixing of the blood and NycoprepTM.


Red blood cells and granulocytes are sedimented to the bottom of the tube while the white blood cells migrate to the interface between the plasma and NycoprepTM layers. They are readily identified as a discrete white band located above the plastic insert.

 

 

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