Separating Solutions Pre-Filled
Pancoll separating solutions from PAN-Seratech are made from a neutral, highly cross-linked, hydrophilic polymer of sucrose with an average molecular weight of 400,000 daltons. Pancoll is suited for separation of lymphocytes and other cell types. The ready-to-use solutions are available in 500 ml bottles (see page 105) as well as in pre-filled ready-to-use tubes with a separating membrane.
|Pancoll human, density 1.077 g/ml||25 x 50 ml
50 x 10 ml
|Pancoll animal, density 1.077 g/ml||50 x 10 ml||ST04-63225|
Pancoll is stable for 36 months at 2 °C to 20 °C. Protect from light!
For lymphocyte separation blood is used which has been defibrinated or treated with anticoagulants (Heparin, EDTA, Citrate), and which is diluted with the same volume of a physiological saline solution. Then the Pancoll solution is carefully covered with a layer of diluted blood in a centrifuge vial, without mixing the phases. After a short centrifugation step (e.g. 800-1000x g for 20-30 minutes without brake) at room temperature the lymphocytes, together with monocytes and platelets, can be harvested from the white blood cells layer between the plasma sample layer and the Pancoll. The separated cells are then washed twice in physiological saline solution to purify the lymphocytes by removing platelets. During centrifugation the cells of the blood sample migrate to the Pancoll layer where they get into contact with the polysaccharide contained in Pancoll. The red blood cells are aggregated by this substance at room temperature immediately. Aggregation causes an increase of the sedimentation rate of the red blood cells which aggregate together with the granulocytes as a sediment at the bottom of the centrifuge vial. Lymphocytes, monocytes and platelets are not so dense and can not enter and pass through the Pancoll layer. These cells are concentrated as white blood cell layer above the Pancoll layer and therefore can be harvested easily by careful pipetting. In subsequent centrifugation steps the lymphocytes are washed to remove remaining platelets, serum and Pancoll. As a result of this process a highly purified suspension of viable lymphocytes and monocytes (PBMC) is obtained.