Isolation of human mononuclear cells
Mononuclear cells (monocytes and lymphocytes) have a lower buoyant density than the erythrocytes and the polymorphonuclear (PMN) leukocytes (granulocytes). The vast majority of mononuclear cells have densities below 1.077 g/ml. These cells can therefore be isolated by centrifugation on an isoosmotic medium with a density of 1.077 g/ml, which allows the erythrocytes and the PMNs to sediment through the medium while retaining the mononuclear cells at the sample/medium interface.
The described method is rapid, simple and reliable and gives excellent results with blood samples from normal individuals and patients.
The contamination of erythrocytes in the mononuclear cell suspension is usually between 3-10 % of the total cell number.
Some immature PMNs may band with the lymphocytes during intense immunosuppressive therapy.
When heparinised blood is used, it is essential to remove most of the platelets, in order to avoid inhibition in the cytotoxicity test.
For every batch produced a Certificate of Analysis showing the actual values of density, osmolality and endotoxins is made available at „diagnostic.serumwerk.com“. We also claim sterility according to Ph.Eur.
Lymphoprep is manufactured, packed and released in compliance with GMP and ISO13485.
Lymphoprep has the same specifications as the expensive PLUS or PREMIUM media from other manufacturers.
Lymphoprep can be used for the preparation of pure lymphocyte suspensions for tissue typing, antilymphocyte sera and immunological research. Thorsby and Brattelie used this technique with only slight modifications in the preparation of pure lymphocyte suspensions for cytotoxicity tests and lymphocyte cultures.
Bøyum, A. (1968)
Separation of leucocytes from blood and bone marrow Scand. J. Clin. Lab. Invest., 21, suppl.97
tested solution with the following specifications
|Sodium diatrizoate||9.1% (w/v)|
|Density||1.077 ± 0.001 g/ml|
|Osmolality||290 ± 15 mOsm|
|Endotoxins||< 1.0 IU/ml|
a sterile solution in the following package sizes:
|Prod. No. 1856||1 x 250 ml|
|Prod. No. 1858||1 x 500 ml|
|Prod. No. 1856||10 x 250 ml|
|Prod. No. 1858||10 x 500 ml|