Characterization of BioSafe SEPAX Manufactured Stem Cell Intended for Cardiac Cell Therapy.

Numerous cardiology groups are actively pursuing the use of hematopoietic stem cells for myocardial delivery. Most published studies have used a manual ficoll density separation procedure to enrich the Mononuclear Cell (MNC) population thus enriching the stem cell content in the final therapeutic cell product. Manual ficoll procedures have certain intrinsic limitations including technologist variability, training time, and risk of contamination due to its open nature. Most open and manual cell processing procedures have been supplanted by recent technology developments in automated and closed devices. The BioSafe SEPAX is a closed system-automated ficoll density separation device that currently has FDA 510k clearance for the processing of umbilical cord blood products. In support of multi-center clinical studies being conducted by the NHLBI Cardiovascular Cell Therapy Research Network (CCTRN), the MD Anderson Cell Therapy Laboratory has evaluated bone marrow derived, MNC enriched, stem cells for their cardiovascular therapeutic potential. Traditional transplantation immunophenotyping was performed on these cells to evaluate hematopoietic stem cells (CD34+ and CD133+), T-Cell subsets, B-Cells, and NK-Cells. Colony Forming Units (CFU) as wells as surrogates used in cardiac regeneration studies including endothelial invasion assays and CXCR4 immunophenotyping (a chemokine receptor involved in cell migration) were also performed. Our data suggests the cells produced using the automated ficoll BioSafe SEPAX device are equivalent to cells produced using a manual ficoll procedure with respect to TNC, MNC, viability, and CD34+ recoveries. Cell subsets and function including invasion and CXCR4 analysis were also similar between the two procedures. The automated method produced a product with statistically higher CFU potential than the manual method suggesting the SEPAX isolates a more potent subset of the mononuclear fraction. These studies support the use of the SEPAX device as for enriching BM-MNCs for the CCTRN. Additional benefits might include the use of a closed system, ease of training, speed of cell manufacturing, and product consistency.