Premium
Density gradient separation of peripheral blood stem cells: comparison of an automated cell processing device and manual methods
Author(s) -
Hillyer C.D.,
Tiegerman K.O.,
Berkman Eugene M.
Publication year - 1990
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.1990.30991048789.x
Subject(s) - leukapheresis , ficoll , peripheral blood mononuclear cell , stem cell , apheresis , hematocrit , chromatography , andrology , medicine , immunology , platelet , biomedical engineering , biology , cd34 , chemistry , biochemistry , microbiology and biotechnology , in vitro
Peripheral blood stem cells were collected from normal donors by leukapheresis on a cell separator. The leukapheresis product contained 1.5 × 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 × 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.