Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy

• Published in: The journal of gene medicine 2005-07, Vol.7 (7), p.926-935
• Main Authors: Hasson, E., Slovatizky, Y., Shimoni, Y., Falk, H., Panet, A., Mitrani, E.
• Format: Article
• Language: English
• Subjects: adeno virus; Animals; beta-Galactosidase - analysis; Colon; Dependovirus - genetics; epithelial-mesenchymal interactions; ex vivo gene therapy; Female; Gene therapy; Gene Transfer Techniques; Genes, Reporter; Genetic Therapy; Genetic Vectors; herpes simplex type-1; Herpesvirus 1, Human - genetics; Leukemia Virus, Murine - genetics; Lung; Male; Mice; Murine leukemia virus; Organ Culture Techniques; Skin; Spleen; Tissue Distribution; Transduction, Genetic; Transplantation, Homologous; Vaccinia virus; Vaccinia virus - genetics
• ISSN: 1099-498X; 1521-2254
• DOI: 10.1002/jgm.740

Background Organ fragments can be cultured for weeks in vitro if they are prepared of microscopic thickness and if the basic organ structure is preserved. Such organ fragments, which we termed micro‐organs (MOs), express in culture endogenous tissue‐specific gene products. We have exploited this methodology to engineer MOs ex vivo by gene transfer. Methods MOs prepared from spleen, lung, colon and skin were infected using: herpes simplex type‐1, adeno virus, vaccinia virus and murine leukemia virus (MuLV), carrying the reporter gene β‐galactosidase. Results All four viral vectors infected MOs in culture, with adeno infection giving significantly higher values. After optimization, high levels of expression (>15% positive cells), comparable to those obtained with the adeno construct, were also obtained using the MuLV construct both in vitro and after implantation into syngeneic hosts. After implantation, the engineered tissue was found to remain localized, become vascularized, and to express the transduced gene for several months. Conclusions The system can be used to study interactions between viruses and tissues both ex vivo and in vivo. Furthermore, the approach proposes a novel platform for ex vivo gene therapy. Such engineered structures could be used as autologous biological pumps for continuous secretion in vivo of gene products of clinical importance. Copyright © 2005 John Wiley & Sons, Ltd.