Hillyer C D, Tiegerman K O, Berkman E M
Division of Hematology/Oncology, New England Medical Center, Boston, Massachusetts.
Transfusion. 1990 Nov-Dec;30(9):824-7. doi: 10.1046/j.1537-2995.1990.30991048789.x.
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.
通过血细胞分离机从正常供体采集外周血干细胞。白细胞分离产物含有1.5×10¹⁰个单核细胞（MNC），并分为两份，分别进行自动或手动的菲可-泛影葡胺密度梯度分离及后续洗涤。在自动过程中，MNC回收率为85%，血小板含量降低64%，最终血细胞比容（Hct）小于1%。手动分离导致MNC回收率为76%，血小板含量降低79%，最终Hct小于1%。然后将纯化的MNC以每毫升4×10⁵个MNC的浓度置于甲基纤维素培养基中。培养一式四份的1毫升等分试样，并在第14天对集落进行计数和分类。自动和手动菲可-泛影葡胺分离的比较表明，总集落数、红系集落数或粒-巨噬细胞集落数无差异。所使用的细胞处理仪快速、可靠、操作简单，并提供一种无菌产物，其中的祖细胞不受自动菲可-泛影葡胺分离的不利影响。
