A simple and effective method for Dr Arne reported on the isolation of mononuclear cells from human blood Bøyum in 1968. For more than 35 years, a commercial medium known as Lymphoprep™ has been widely used to isolate these cells. Mononuclear cells (monocytes and lymphocytes) have a lower buoyant density than erythrocytes and polymorphonuclear leukocytes (PMN) (granulocytes).
The vast majority of mononuclear cells have densities below 1.077g/ml. Therefore, these cells can be isolated by centrifugation in an isoosmotic medium with a density of 1.077 g/ml, allowing the red blood cells and the PMN to sediment through the m-medium preserving mononuclear cells in interface shows/medium. The method described is fast, simple and reliable and gives excellent results with blood samples from individuals and normal patients.
Contamination of erythrocytes in the mononuclear cell suspension is generally between 3-10% of the total cell phone number. Some immature PMNs can form bands with lymphocytes during intense immunosuppressive therapy. When heparinized blood is used, it is essential to remove most of the platelets, to prevent inhibition in the cytotoxicity test. Lymphoprep ™ is a sterile preparation and tested solution of endotoxins with the following specifications:
- Sodium diatrizoate: 9.1% (w/v)
- Polysaccharide: 5.7% (w/v)
- Density: 1.077 ± 0.001 g/ml
- Osmolality: 290 ± 15 mOsm
- Endotoxins: < 1.0 EU/ml
Each batch of Lymphoprep™ is tested at the level of endotoxins using a speci c LAL test. Our objective is to produce batches with an endotoxin level less than or equal to 0.13 EU/ml. Lymphoprep™ is manufactured, packaged and released in compliance with:
1. Current EU guidance on good manufacturing practice
2. Fresenius Kabi AS Quality System
3. Fresenius Kabi AS manufacturing license
4. Axis-Shield PoC Quality System
Lymphoprep™ has the same specifications as the expensive PLUS or PREMIUM media from other manufacturers. Lymphoprep™ can be used for the preparation of cigars lymphocyte suspensions for tissue typing, antilymphocyte sera and immunological research. Thorsby and Brattelie used this technique with slight modifications in the preparation of suspensions of pure lymphocytes for cytotoxicity tests and lymphocyte cultures.