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Isolation of lymphocytes from whole blood
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Objectives

 

To isolate the lymphocyte from whole blood by density gradient centrifugation method

 

Theory

 

Blood is made up of a suspension of special cells in liquid called plasma. A lymphocyte is small white blood cells, usually 7 to 8 micrometers in length present in the blood. They are the only blood cells which can be transformed into actively proliferating cells. Lymphocytes are originated from lymphatic tissue throughout the body. This lymphocytes collected from blood have been in frequent use in the field of immunology. It helps to protect the body from diseases, invasion of foreign bodies, tumors and infections. For example, lymphocytes are used for the for immunological studies directed to histocompatibility antigens and cellular immunity. Lymphocytes are also used for determining lymphocyte blastotransformation or for determining the proportion of T cells to B cells, and for trying to separate helper T cells and Suppressor T cells from T cells.

 

Typical process for carrying out lymphocyte separation is the density gradient centrifugation. Centrifugation is the commonly used method for processing blood so that the cells and particles of the same size, shape and density sediment as separate zones without convection. Whole blood can be easily and reproducibly fractionated into different components through differential centrifugation and selective removal. The separated blood components can subsequently be used for their respective clinical and scientific applications and investigations. Aqueous density media have been used with centrifugation to separate and isolate different blood cell types. Separation of lymphocytes from whole blood using HISTOPAQUE-1077 is based on a method first described by Boyum1in 1968. The separation medium, HISTOPAQUE-1077, is an aqueous solution of a high molecular weight polysaccharide and sodium diatrizoate, aniodinated nonionic compound, adjusted to a density of 1.077 ± 0.001. This medium facilitates rapid recovery of viable lymphocytes from small volumes of blood.  

 

 

 

 

 

 

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