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Human peripheral blood mononuclear cell subfractionation using counterflow centrifugation elutriation
C Almici1, L Mangoni, C Carlo-Stella
1Cattedra di Ematologia, Università di Parma, Italy.
Haematologica
|March 1, 1991
Summary
Counterflow Centrifugation Elutriation (CCE) effectively separates human peripheral blood cells. This method yields highly purified lymphocyte and monocyte fractions with excellent cell viability and recovery for further research.
Area of Science:
- Cell biology
- Immunology
- Biotechnology
Background:
- Counterflow Centrifugation Elutriation (CCE) separates cells based on sedimentation.
- Evaluating CCE for normal human peripheral blood fractionation is crucial.
Purpose of the Study:
- To assess the efficacy of CCE in fractionating normal human peripheral blood.
- To determine the purity and viability of cell subpopulations obtained via CCE.
Main Methods:
- Human peripheral blood mononuclear cells (PBMC) were fractionated using CCE.
- Cells were separated into three distinct fractions based on elution flow rates.
Main Results:
- Two fractions enriched in lymphocytes (<2% and <4% monocytes) were obtained, responsive to PHA.
- A third fraction predominantly contained monocytes (>85%), non-responsive to PHA.
- High cell recovery (>93%) and viability (>96%) were achieved across all fractions.
Conclusions:
- CCE is a suitable technique for large-scale cell separations.
- CCE provides highly purified cell fractions for subsequent manipulation.
- The method preserves cell viability and function for downstream applications.