P257 Automated isolation of lymphoid (CD3+, CD19+, CD56+) and myeloid (CD33/CD66B+, CD15+) subsets for use in lineage-specific chimerism analysis

Aim Lineage-specific chimerism analysis is used to monitor engraftment in different cell subsets following hematopoietic cell transplantation. Isolation of specific cell types can be time consuming, and so methods that automate the process are of great value to a busy chimerism laboratory. We have recently developed a method (EasySep™) to rapidly isolate different lymphoid (CD3+, CD19+, CD56+) and myeloid (CD33/CD66b+, CD15+) subsets from whole blood or buffy coat (BC). The aim of this study was to compare cell isolation performance for all 5 subsets between the automated RoboSep™-S and RoboSep™-16 instruments, which isolate 4 and 16 samples, respectively, and the manual silver “The Big Easy” EasySep™ magnet. Methods CD3+, CD19+, CD56+, myeloid and CD15+ cells were isolated from 0.9 mL BC samples from 5 different donors on each platform as per the manufacturer’s instructions. Following cell isolation, the percent purity by flow cytometry, the average number of cells isolated from the 0.9 mL BC sample, and the ug of DNA obtained from 2.5 × 10e5 isolated cells using QIAGEN spin columns were determined for each cell isolation platform (mean ± SD). Results Download high-res image (525KB) Download full-size image Conclusions No significant difference in cell purity, number of cells acquired, or ug of DNA obtained between the manual or automated isolation platforms was identified for any of the cell subsets analyzed (ANOVA). Thus use of the automated RoboSep™ platforms to isolate lymphoid and myeloid cell subsets will save time by increasing sample throughput up to 16-fold for busy chimerism testing laboratories.