Isolation of human mononuclear cells – easy handling
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 success of the standard method for isolation mononuclear cells using Lymphoprep depends to a large extent on the careful layering of the diluted blood sample on top of the centrifugation medium to maintain a sharp interface between the two layers. This procedure requires some practise and can be time consuming with large numbers of samples.
Lymphoprep Tube is a sterile tube in which the Lymphoprep is contained below a plastic filter disc. This allows diluted blood to be poured simply and directly into the tube, the disc preventing any mixing with the separation medium.
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 Tube can be used for the preparation of pure lymphocyte suspensions for tissue typing, antilymphocyte sera and immunological research. Thorsby and Bratlie used this technique with only slight modifications in the preparation of pure lymphocyte suspensions for cytotoxicity tests and lymphocyte cultures.
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.
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. 18002||30 x 15 ml (total tube size)
conical centrifuge tubes
(each tube filled with 2 ml Lymphoprep)
|Prod. No. 18001|| 18 x 50 ml (total tube size)
conical centrifuge tubes
(each tube filled with 10 ml Lymphoprep)