TY - JOUR
T1 - Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy
AU - Hasson, E.
AU - Slovatizky, Y.
AU - Shimoni, Y.
AU - Falk, H.
AU - Panet, A.
AU - Mitrani, E.
PY - 2005/7
Y1 - 2005/7
N2 - 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.
AB - 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.
KW - Adeno virus
KW - Epithelial-mesenchymal interactions
KW - Ex vivo gene therapy
KW - Herpes simplex type-1
KW - Murine leukemia virus
KW - Vaccinia virus
UR - http://www.scopus.com/inward/record.url?scp=22544475353&partnerID=8YFLogxK
U2 - 10.1002/jgm.740
DO - 10.1002/jgm.740
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C2 - 15744776
AN - SCOPUS:22544475353
SN - 1099-498X
VL - 7
SP - 926
EP - 935
JO - Journal of Gene Medicine
JF - Journal of Gene Medicine
IS - 7
ER -