Enrichment of Mononuclear Cells From Cryopreserved Cord Blood Units Using the Purecell” Select System

• Published in: Blood 2009-11, Vol.114 (22), p.2159-2159
• Main Authors: Spencer, Bryan P, Kaur, Indreshpal, Fong, Patrick, Karandish, Safa, Ponweera, Nirmali, Baez, Omar, Shpall, Elizabeth J, Champlin, Richard E, McMannis, John D
• Format: Article
• Language: English
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V114.22.2159.2159

Abstract 2159 Poster Board II-136 Many procedures for manufacturing clinical products for cellular therapy involve enrichment of mononuclear cells (MNC). The most common procedure is density gradient separation. Disadvantages of this procedure are low yield of cells especially with cryopreserved products and the open events during processing. We evaluated use of the Purecell” Select System (PALL Medical, NY) for enriching MNC from cryopreserved Cord Blood Units (CBU). Initial experiments were performed to optimize the system for recoveries of total nucleated cells (TNC), MNC, neutrophils, lymphocytes, monocytes and CD34+/CD3+ cells. We evaluated thaw/predilute/filter vs thaw/filter, starting volumes (30 — 95mls) and three different methods for harvesting cells from the filter (standard method, input bag rinse and harvest port rinse). Once conditions were optimized, cryopreserved CBU were thawed and split into two fractions. One half of the product was diluted and processed on the Purecell” Select System. The other half was washed and ficolled. The MNC fraction was CD3+ enriched using CD3/28 beads and then cultured with rIL-2 (200 units/ml) for 14 days. The absolute number of CD3+ cells post culture, fold expansion and viabilities of these cells were determined. The Purecell” procedure was 15 times faster than the ficoll method. Optimal volume to load onto the filter was 50ml. When MNC were harvested by the three recommended procedures, there was no difference in recoveries of TNC, MNC, CD34+ and CD3+ cells and neutrophils. However, the lymphocyte and monocyte recoveries were higher (p