Density gradient separation of peripheral blood stem cells: comparison of an automated cell processing device and manual methods.
Peripheral blood stem cells were collected from normal donors by leukapheresis on a cell separator. The leukapheresis product contained 1.5 x 10(10) mononuclear cells (MNCs) and was divided into two aliquots that underwent either automated or manual density gradient separation with ficoll-hypaque and subsequent washing. In the automated process, recovery of MNCs was 85 percent, reduction in platelet content was 64 percent, and the final hematocrit (Hct) was less than 1 percent. The manual separation resulted in 76-percent MNC recovery, a 79-percent reduction in platelet content, and a final Hct of less than 1 percent. The purified MNCs were then placed in methylcellulose culture at a concentration of 4 x 10(5) MNCs per mL. Quadruplicate 1-mL aliquots were cultured, and colonies were counted and classified on Day 14. Comparison of automated and manual ficoll-hypaque separations demonstrated no differences in the total, erythroid, or granulocyte-macrophage colony numbers. The cell processor used is fast, reliable, uncomplicated, and provides a sterile product containing progenitor cells that are not adversely affected by the automated ficoll-hypaque separation.
Hillyer
C D
CD
Division of Hematology/Oncology, New England Medical Center, Boston, Massachusetts.
Tiegerman
K O
KO
Berkman
E M
EM
D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D006801:Humans; D007937:Leukapheresis / Q000379:methods; D013234:Stem Cells / Q000166:cytology*