A rapid and simple method for the separation of pure lymphocytes from horse blood

A method for the separation of pure and viable lymphocytes and granulocytes from the same blood sample in horses was reported. By centrifuging equine heparinized blood at 100× g for 10 min at room temperature (r.t.), the resulting supernatant plasma was an almost pure (97.71±0.30%; n=15) suspension of highly viable (98.72±0.28%) lymphocytes. When sodium citrate was used as an anticoagulant, lymphocyte suspensions collected in the same manner showed lower purity (87.89±1.59%; n=9) and higher yields (56.56±3.89%, n=9 versus 36.11±2.23%, n=15). Where needed, a further centrifugation at 250× g for 3 min (r.t.) of heparinized lymphocyte preparations removed an average of 87.39% ( n=15) contaminating platelets. A suspension of 85.96±2.20% pure granulocytes (93.23±1.74% neutrophils; n=14) with minimal contamination by erythrocytes and high viability (93.11±1.26%) was obtained by performing a flash red blood cell lysis on the white-greyish layer resulting from the centrifugation of the heparinized blood samples. Among the several methods available, the procedure described herein is easy, rapid, cheap and reproducible.