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Furthermore, we established a stable, human alpharetroviral packaging cell line in which we deleted the entry receptor (SLC1A5) for RD114/TR-pseudotyped ARVs to prevent superinfection and enhance genomic integrity of the packaging cell line and viral particles.
We propose here that a MSC packaging cell line could be employed for in situ gene therapy of solid tumors.
The viral packaging cell line 293FT was from Invitrogen.
First, the health of the 293T packaging cell is critical.
The 293T-based Pheonix ecotropic packaging cell line (provided by G. Nolan, Stanford Univ).
The φNX packaging cell line (Orbigen) was transfected using Lipofectamine 2000 (Invitrogen).
Stable cell lines were generated by transfecting the 293FT packaging cell line.
The Phoenix A packaging cell line was maintained as recommended by the ATCC.
RV vector supernatants were produced by transient transfection of the amphotropic Phoenix packaging cell line.
Each recombinant plasmid was introduced into the 293gpg retrovirus packaging cell line [21] by transient transfection, followed by harvesting.
A PT67 retrovirus packaging cell line was obtained from Clontech, and maintained in DMEM with 10% FBS.
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