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Updated: Dec 7, 2025
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Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
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A method for separation and purification of mouse splenocytes by density gradient centrifugation
Yao Lu1, Chenghao Fu2, Chao Xia1
1College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, P. R. China.
Preparative Biochemistry & Biotechnology
|September 28, 2020
Summary
Optimizing density gradient centrifugation improves spleen cell separation purity. This method effectively isolates lymphocytes and myeloid-derived suppressor cells (MDSCs) from tumor-bearing mouse spleens, aiding cancer research.
Area of Science:
- Immunology
- Cell Biology
Background:
- The spleen is a crucial peripheral lymphoid organ with complex immunocyte composition.
- Understanding immunocyte behavior requires isolating specific subpopulations without heterogeneity.
- Density gradient centrifugation offers high throughput but suffers from low purity.
Purpose of the Study:
- To optimize density gradient centrifugation for separating mouse splenocytes.
- To improve the purity of isolated lymphocytes and myeloid-derived suppressor cells (MDSCs).
Main Methods:
- Density gradient centrifugation was optimized using varying density gradients for tumor-bearing mouse splenocytes.
- Cell layers were analyzed for purity and gene expression of chemokine receptors.
- The method was validated on healthy mouse splenocytes.
Main Results:
- Optimized separation yielded 84.9% lymphocytes in layer 2, 54.2% monocytic-like MDSCs (Mo-MDSCs) in layer 4, and 85.5% polymorphonuclear MDSCs (PMN-MDSCs) in layer 6.
- Gene expression patterns confirmed cell identities.
- The method successfully separated splenocytes from both tumor-bearing and healthy mice.
Conclusions:
- Optimized density gradient centrifugation enhances splenocyte separation purity.
- This technique provides a foundation for studying immunocyte roles in cancer development.
- Improved isolation of specific cell subpopulations is vital for immune response research.
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