MSC Separation on Bioactive Molecule-Immobilized Column

Mesenchymal stem cell (MSC) is the ideal cell source in regenerative medicine due to its multipotent and proliferative activity which are essential for treating the defective tissues. Separation and purification of MSCs from living body are critical step for autologous cell plantation and tissue engineering. FACS or Ficoll methods were generally selected for separating the MSCs, but the isolated cells are not homogeneous. Moreover, these methods have the disadvantage to require some reagents which allow the dissociation of the cells and the antibodies, and the cell damage during the purification procedure is another problem. The therapeutic effects of target tissue using MSCs were reported in clinical research, but these contaminations may result in unknown side effects. To secure the safety of MSC plantation as a clinical treatment, it is necessary to develop novel procedure for separating the homogeneous cells. In this research, we develop a cell separation column that was immobilized with antibody against MSC. The cells injected to the column roll on the inner surface of the column under a flow condition like a rolling adhesion phenomenon of the leukocyte in the blood vessel [1]. Polyacrylic acid was grafted on the surface of the polyethylene or silicone tube by ozone-induced graft polymerization. The anti-human CD34 antibody was immobilized on the surface. The cell suspension of KG-1a (CD34 positive) or HL-60 (CD34 negative) passed through the column, and the number and surface marker pattern of the cells in each elution fraction were analyzed by FACS system. The MSCs derived from mouse bone marrow was also injected in the column and the number and pattern was evaluated by FACS.