[Frontiers in Bioscience 7, d752-764, March 1, 2002]

RETROVIRAL PACKAGING CELLS ENCAPSULATED IN THERACYTE IMMUNOISOLATION DEVICES ENABLE LONG-TERM IN VIVO GENE DELIVERY

Anna Krupetsky, Zahida Parveen, Elena Marusich, Adrienne Goodrich, and Ralph Dornburg

Thomas Jefferson University, Division of Infectious Diseases, 1020 Locust Street, Suite 329, Philadelphia, PA

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Material and Methods
3.1. Cells
3.2. Radiation of helper cells
3.3. TheraCyteª preparation and loading
3.4. In vitro infection via TheraCyteª
3.5. FACS analysis
3.6. Immunostaining
3.7. Test for RCR
4. Results
4.1. In vitro experimental system
4.2. In vivo studies
5. Discussion
6. Acknowledgement
7. References

1. ABSTRACT

The method of delivering a therapeutic gene into a patient is still one of the major obstacles towards successful human gene therapy. Here we describe a novel gene delivery approach using TheraCyte immunoisolation devices. Retroviral vector producing cells, derived from the avian retrovirus spleen necrosis virus, SNV, were encapsulated in TheraCyte devices and tested for the release of retroviral vectors. In vitro experiments show that such devices release infectious retroviral vectors into the tissue culture medium for up to 4 months. When such devices were implanted subcutaneously in SCID mice, infectious virus was released into the blood stream. There, the vectors were transported to and infected tumors, which had been induced by subcutaneous injection of tissue culture cells. Thus, this novel concept of a continuous, long-term gene delivery may constitute an attractive approach for future in vivo human gene therapy.