Introduction
Lymphocytes are essential immune system cells that fight infection. Isolating them from blood is necessary for various immunological studies. Lymphocyte separation media (LSM) is a solution used in density gradient centrifugation, a common technique to enrich lymphocytes from whole blood samples.
LSM Function
LSM typically contains ficoll, a large sugar molecule, and hypaque, a contrast agent. These create a density gradient in the solution. Blood anticoagulated with a specific substance (e.g., sodium citrate, heparin) is layered on top of the LSM in a centrifuge tube. Centrifugation separates the blood components based on their weight.
Lymphocyte Enrichment
Red blood cells (RBCs), being the heaviest, sink to the bottom. Plasma, the liquid part of blood, forms the top layer. Lymphocytes, monocytes, and granulocytes (all considered peripheral blood mononuclear cells (PBMCs) due to having one nucleus) have a medium density and form a band between the plasma and the LSM. The LSM is designed to position this band clearly, allowing researchers to collect the PBMCs easily.
Types of LSM
Several commercially available LSM options exist, with slight variations in ingredients and density. These variations can affect how well the separation works and the yield of specific PBMC types (e.g., T cells, B cells). Researchers should choose an LSM formulation optimized for their specific experiment.
Considerations
- Cell Purity: Isolated PBMCs may still contain some granulocytes and erythrocytes. Additional purification steps might be necessary depending on the experiment.
- Cell Viability: Careful handling of blood samples and following manufacturer instructions are crucial for maintaining high lymphocyte viability.
- Anticoagulant Choice: The type of anticoagulant used can influence cell viability and downstream applications. Researchers should select an anticoagulant compatible with their specific needs.
Review about the BioSci™ Human Lymphocyte Separation
Medium:
Lymphocytes are essential immune system cells that fight infection. Isolating them from blood is necessary for various immunological studies. Human Lymphocyte Separation Media (LSM) is a solution used in density gradient centrifugation, a common technique to enrich lymphocytes from whole blood samples.
lt is suitable for separating human peripheral blood lymphocytes as well as separating mononuclear cells of most mammals.
Features
1. Convenient
• The operation is simple.
2. Safe
• Il is sterile and has low toxicity.
3. Excellent Separation Performance
• Il sustains high cell viability, high purity and high yield.
• Layers are obvious, strips are clear, and particles are better than the national standard.
Conclusion
LSM is an important tool for isolating lymphocytes from human blood. Its efficiency, reproducibility, and ease of use make it popular for various immunological studies. However, researchers should consider potential limitations, such as cell purity and the impact of anticoagulant choice, to ensure optimal results for their specific applications