About Polymorphprep ™
Polymorphprep ™ is an endotoxin-tested, sterile, prepared solution used to isolate polymorphonuclear granulocytes from blood. It is the ideal medium for neutrophils and eosinophils. Mononuclear and polymorphonuclear leukocytes separate into two distinct bands free of red blood cells. The temperature of the blood sample and the medium should be kept between 18-22 ° C. Variations in temperature can affect the density and viscosity of the Polymorphprep ™ solution, which can lead to a poor result.
The method is effective only with undiluted whole blood, not with a fraction rich in leukocytes.
Storage Conditions and Stability of Polymorphprep ™
• Polymorphprep ™ should be stored at room temperature (20 ° C).
• It is stable for 3 years as long as the solution is kept sterile and protected from light.
• Prolonged exposure to direct sunlight leads to the release of iodine from the sodium diatrizoate molecule (this effect is negligible when working with the solution on a day-to-day basis).
Isolation of human polymorphonuclear cells
The solution contains sodium diatrizoate and dextran 500 in the following concentrations:
• Sodium diatrizoate 13.8% (p / v)
• Dextran 500 8.0 (p / v)
How Polymorphprep ™ Works
Polymorphonuclear leukocytes (PMNs) have a lower float density than erythrocytes (1.09-1.11 g / ml) but a higher float density than mononuclear cells (more than 1.085 g / ml). Therefore, it is difficult to isolate polymorphonuclear granulocytes using a barrier of density similar to that used for mononuclear cells. Various procedures have been developed to overcome these problems. The high osmolality of Polymorphprep ™ causes erythrocytes to lose water and shrink, thus increasing their effective buoyancy density.
This allows the dextran-aggregated erythrocytes to sediment rapidly through the dense medium. As the cells settle further in the medium, the osmotic gradient between the medium and the erythrocytes decreases. This means that the water loss from the erythrocytes is greatest at the top of the Polymorphprep ™ and progressively decreases as they settle further. A diatrizoate gradient forms within the density barrier. The PMN band within this density gradient, while mononuclear cells remain at the sample/medium interface.
Post mentioning Polymorphprep ™
1. Ferrante, A. and Thong, Y.H. (1980): Optimal conditions for the simultaneous purification of mononuclear and polymorphonuclear leukocytes from human peripheral blood by the Ficoll-Hypaque method. J. Immunol. Methods 36,109
2. Sofia Tedelind and al. (2007) Anti-inflammatory properties of short-chain fatty acid acetate and propionate: a study with relevance for inflammatory bowel disease
3. Iwaniuk A, Jab? OR? Ska E, Jab? OR? Ski J, Ratajczak-Wrona W, Garley M. 2015. Expression of proteins selected from the extrinsic and intrinsic pathways of apoptosis in human leukocytes exposed to N-nitrosodimethylamine. Hum Exp Toxicol. 6: 591–600