Related Experiment Videos
Comparative study of eosinophil purification on Nycodenz, Metrizamide and Percoll density gradients
N W Brattig1, C E Medina-De la Garza, F W Tischendorf
1Department of Clinical Chemistry, Bernhard-Nocht-Institut für Schiffs- und Tropenkrankheiten, Hamburg, FRG.
European Journal of Haematology
|August 1, 1987
Summary
Purifying human eosinophils for research is challenging. This study introduces Nycodenz for eosinophil isolation, comparing it with Percoll and metrizamide, and evaluating cell yield, purity, and function.
Area of Science:
- Immunology
- Cell Biology
- Parasitology
Background:
- Human eosinophil isolation for metabolic and parasitic interaction studies is difficult due to purity and quantity challenges.
- Effective purification methods are crucial for advancing research on eosinophil functions.
Purpose of the Study:
- To introduce and evaluate Nycodenz density gradient centrifugation for human eosinophil purification from peripheral blood.
- To compare Nycodenz with established methods like Percoll and metrizamide gradient centrifugation.
- To assess cell yield, purity, and functional integrity of isolated eosinophils.
Main Methods:
- Density gradient centrifugation using Nycodenz, Percoll, and metrizamide.
- Comparison of purification efficiency based on cell yield and purity.
- Assessment of functional integrity through metabolic assays (superoxide anion generation) and phagocytosis assays.
- Evaluation of eosinophil interaction with helminthic larvae (Onchocerca volvulus).
Main Results:
- Percoll gradients yielded the purest eosinophil preparations (94% +/- 15%) compared to Nycodenz (58% +/- 29%) and metrizamide (88% +/- 13%).
- Nycodenz centrifugation resulted in low eosinophil recovery rates (29% +/- 19%) compared to metrizamide (60% +/- 21%) and Percoll (70% +/- 16%).
- Eosinophils isolated by all three methods maintained metabolic function and phagocytic capabilities, with Percoll-isolated cells adhering to and immobilizing Onchocerca volvulus larvae.
Conclusions:
- While Nycodenz can be used for eosinophil purification, Percoll offers superior purity for eosinophils from blood with moderate eosinophilia.
- All tested methods preserve eosinophil functional integrity, including metabolic activity and phagocytosis.
- Percoll-isolated eosinophils demonstrate functional interaction with helminthic parasites, highlighting their utility in antiparasitic research.