272. Development of |[ldquo]|Full|[rdquo]|-Closed Sterile and Automatic Retroviral Gene Transfer System for Clinical Gene Therapy

Manipulating large number of cultured cells for gene therapy has risk of microbiological contamination constantly. It would become more serious under the situation like re-infusion of such cells to the compromised host. Our clinical protocol, HSV-TK gene transfer into donor lymphocytes for control of allogeneic Graft-versus-leukemia and Graft-versus-host-disease (Japanese trial), is one of such sensitive protocol by meaning of immunological conditions of patients and manipulation steps. To avoid those problems we are developing manipulation protocol for retroviral gene transfer to human T lymphocytes using automatic cell processor CytoMateTM, magnetic cell selection system IsolexTM and CO2 permeable cell culture bag OpticytoTM. All of manipulations, except centrifugation for gene transfer, had been performed in "full"-closed sterile conditions. Function and sterility of final products, HSV-TK gene transferred lymphocytes, were evaluated by comparing with "non"-closed system, using centrifugation tubes and flasks.