115-P: AUTOMATED HIGH THROUGHPUT ISOLATION OF LYMPHOID AND MYELOID CELLS DIRECTLY FROM WHOLE BLOOD USING RoboSep-HT

Abstract

Aim Process automation is an important requirement for streamlining and standardizing technical procedures, such as cell isolation, in a routine laboratory. We have previously developed RoboSep™, the first instrument to fully automate the cell labeling and separation steps of an immunomagnetic cell isolation procedure. RoboSep™ uses EasySep™ reagents and protocols and can isolate up to 4 individual samples at one time. More recently we have developed a high throughput version of RoboSep™, called RoboSep-HT, which can isolate 16 samples from up to 4 different cell types in about 1 hour. Like RoboSep™, RoboSep-HT uses disposable filter tips to avoid cross contamination between samples and is operated using an intuitive graphical user interface. Sample barcoding is a standard feature. In this study, positive selection of CD3+ and myeloid+ cells directly from whole blood using EasySep™ reagents was performed on the new RoboSep-HT and compared to performance using the classic RoboSep™. Methods 1mL of fresh, 24-hour or 48-hour old whole blood was diluted with an equal volume of 1xRBC lysis buffer and loaded on each instrument. Purity (flow cytometry), recovery (cell counts) and DNA yield (OD) were assessed following isolation using each robot. Replicates were averaged prior to statistical analysis (t-tests). Results There was no significant difference in either purities, cell recoveries or DNA yields when CD3 and myeloid cell isolation was performed on the RoboSep-HT versus the classic RoboSep™. [ Table 1 ] Conclusions RoboSep-HT can be used for the high throughput isolation of multiple cell types and is ideally suited for cell isolation prior to chimerism testing. McQueen: STEMCELL Technologies: Employee. Kellerman: STEMCELL Technologies: Employee. Frehlick: STEMCELL Technologies: Employee. Gromadzki: STEMCELL Technologies: Employee. Poon: STEMCELL Technologies: Employee. Woodside: STEMCELL Technologies: Employee. Thomas: STEMCELL Technologies: Employee.