CONSISTENT AND HIGH-RATES OF GENE-TRANSFER CAN BE OBTAINED USING FLOW-THROUGH TRANSDUCTION OVER A WIDE-RANGE OF RETROVIRAL TITERS

Flow-through transduction methods have been developed to overcome phys ical limitations imposed by Brownian motion on retroviral delivery. Th is method uses net fluid flow of retroviral supernatants through a por ous membrane on which the target cells are placed. It is shown that in comparison to static transduction methods, flow-through transductions have the following advantages: (i) flow-through transductions lead to transduction rates that exceed those obtained by static transduction; (ii) flow-through transductions lead to high transduction rates even at low viral concentrations, eliminating many of the concerns associat ed with the production of high-titer virus supernatants; (iii) flow-th rough transductions are insensitive to viral titers, eliminating the n eed to produce consistently retroviral supernatants at given virus con centrations; (iv) flow-through transductions can be carried out withou t the use of polycations, such as polybrene; and (v) the volume of vir al supernatants needed for gene transfer can be sharply reduced. Taken together, these advantages of flow-through transductions are likely t o lead to their widespread use for gene transfer work, both in researc h and clinical settings.