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Cellular genic therapy: evaluation of mesenchymal stem cells engraftment transduced by an Ecdysone inductible lentiviral vector expressing luciferase gene
Author(s) -
NOCHEZ Y,
COLLIN C,
BELLICAUD D,
CARDON A,
GICQUEL JJ,
PISELLA PJ,
GABISON E,
PAGES JC
Publication year - 2010
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2010.220.x
Subject(s) - luciferase , mesenchymal stem cell , biology , transgene , genetic enhancement , viral vector , gene delivery , microbiology and biotechnology , transfection , gene , genetics , recombinant dna
Purpose Gene therapy might offer an opportunity to improve new treatments in ophthalmology. Several studies, using limbal stem cells or mesenchymal stem cells (MCSs), have confirmed the usefulness of these cells to repair epithelial and stromal corneal tissue. Methods We designed a strategy aimed at expressing Luciferase gene. We have tested the viability of mesenchymal stem cells modified by a lentiviral vector delivery system. To allow an adaptative treatment, the target genes would be transcriptionally controlled by the use of an Ecdysone inductible promoter. Results Construction of a lentiviral plasmid with the transgene (luciferase) and inductible promoter: Ecdysone by recombination. Construction of a lentiviral plasmid with the RXR gene and VgEcR gene and a constitutive enhancer. Transfection of 293 FT cells for lentiviral production. MSCs Transduction by “V/R” lentivirus and by “E/Transgene” lentirus. We have tested induction of Ecdysone system and transcription of the transgene (luciferase) in MSCs adding Ponasterone. Conclusion The expression of luciferase, using conditionnal expression vectors, could be applied after an engraftment of mesenchymal stem cells for pathological wound healing in Sprague‐Dawley rat cornea.