Lymphocyte Separation via Density Gradient Centrifugation: A Reliable Technique for Researchers
Peripheral blood mononuclear cells (PBMCs), encompassing lymphocytes and monocytes, play a pivotal role in immunological investigations. Density gradient centrifugation (DGC) is a well-established technique for isolating PBMCs from whole blood, owing to the distinct buoyant densities of various blood cell populations. This article delves into the principles and practical considerations of DGC for researchers seeking a reliable method for lymphocyte enrichment.
Theoretical Underpinnings
DGC exploits the inherent differences in cell density. Lymphocytes and monocytes (1.075-1.090 g/mL) possess lower densities compared to red blood cells (RBCs) and granulocytes (around 1.092 g/mL) [1]. Platelets, the lightest components (1.030-1.035 g/mL), typically form a top layer. A density gradient medium, such as Ficoll-Paque PLUS, creates a stratified interface within the centrifuge tube. Following centrifugation, RBCs and granulocytes sediment due to their higher density, while PBMCs accumulate at the plasma-medium interface.
Advantages of DGC
- Rapidity and Simplicity: DGC offers a swift and straightforward approach for PBMC isolation, streamlining downstream applications.
- Minimal Cytotoxicity: The separation medium exhibits minimal cytotoxicity towards lymphocytes, ensuring the viability of isolated cells for functional assays.
- High Cell Yield: DGC effectively enriches PBMCs, providing a substantial number of lymphocytes for further experimentation.
Optimizing DGC for Research
- Temperature Control: Maintaining the separation medium at room temperature (20 ± 5°C) prior to use is crucial. Colder temperatures can negatively impact separation efficiency.
- Blood Sample Processing: Freshly collected blood is recommended for optimal results. Extended storage can lead to increased erythrocyte contamination in the isolated PBMC fraction.
- Dilution Considerations: Dilation of the blood sample with an isotonic solution (e.g., phosphate-buffered saline or 0.9% NaCl) can be employed to minimize RBC aggregation and enhance lymphocyte yield.
Supplier Proposition
at Maxanim Inc. , they understand the critical role of reliable lymphocyte isolation in your research endeavors. We offer high-quality, pre-sterilized Lymphocyte Separation Medium, formulated for optimal PBMC enrichment. Our medium ensures:
- Consistent Performance: Experience superior separation efficiency with every experiment.
- Preserved Cell Viability: Isolate viable lymphocytes for downstream functional assays.
- Convenient Storage: Store the medium at 2-8°C for long-term shelf life.
All Dakewe products for lymphocyte separation among other products are now available in Gentaur group, thanks to a strong partnership.
Contact us today to learn more about our lymphocyte separation solutions and discuss how we can empower your research!
- Biosci™ Lymphocyte Separation Tube for Human Peripheral Blood
- Biosci™ Human Lymphocyte Separation Medium
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Biosci™ Mouse Lymphocyte Separation Medium