Pall’s Purecell Select System is an alternative technology to density gradient centrifugation for the enrichment of MNC from whole blood or cord blood. Many density separation media are available commercially, of which Ficoll-Paque* is the most widely used.
Pall’s filtration-based system provides researchers with an alternative enrichment technology that is significantly faster than traditional methods and results in a higher cell yield than Ficoll separations. The MNC recovery from the Purecell Select System typically averages > 70% (Figure 1). MNC recovery across the WB volume range processed by the Pall system (20 to 120 mL) typically averages > 65% (data not shown).
Percent Recovery of Major Leukocyte Populations
Percent recovery of major cell types in 50 mL WB starting material, Purecell Select System processed samples and Ficoll samples. Percent recovery is calculated based on WBC, platelet and red blood cell (RBC) concentration (Cell-Dynu 1800); percent lymphocytes, monocytes and granulocytes is based on phenotype data (BD FACSCalibur system). Duplicates are averaged. Means +/- 1 SD from 6 donors are reported.
Phenotypic analysis of processed samples shows that the Purecell Select System yields more cells of all major types than are recovered with Ficoll separations (Figure 2). The cell population from the Purecell Select System differs from the cell population recovered from Ficoll in that the cell population from the Pall system has a higher concentration of granulocytes and red blood cells. Researchers will need to assess the specific requirements of their applications. Protocol modifications can reduce the residual red blood cell concentration.
Total Cell Number of Major Leukocyte Populations
Total number of major cell types in 50 mL WB and processed samples. Phenotype data (BD FACSCaliber system) averages from 6 donors +/- 1 SD are reported.
Cell viability at 24 hours using propidium iodide is comparable to cell viability from Ficoll processing.
Viability of WBC Population
|
| WB |
Purecell Select System |
Ficoll |
|
|
|
| 95.7% +/- 1.5 |
96.0% +/- 1.4 |
97.3% +/- 3.2 |
|
Percent viability of white blood cells (WBC) was determined by dye exclusion using propidium iodide. The percent viability was calculated by the percentage of unstained versus total cells as determined by flow cytometry (BD FACSCalibur*system). N=5.
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