Suicide gene therapeutic agent for brain tumors using pluripotent stem cell
A technology of pluripotent stem cells and brain tumors, applied in the direction of artificially induced pluripotent cells, antineoplastic drugs, animal cells, etc., can solve problems such as limited results, insufficient diffusion, and difficult treatment methods
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Embodiment 1
[0107] [Example 1] Suicide gene cell therapy for glioma cells
[0108] (A) Materials and methods
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[0110] The human iPS cells (1210B2) used were obtained from the iPS Cell Research Institute (CiRA), Kyoto University. 1210B2 A method for introducing reprogramming factors into human peripheral blood mononuclear cells by using episomal vectors (Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, Goshima N, Yamanaka S. An efficient nonviral method to generate integration-free human-induced pluripotent stemcells from cord blood and peripheral blood cells. Stem Cells. 2013 Mar; 31(3):458-66.) and constructed. Human iPS cells were seeded on plastic culture dishes coated with iMatrix-511 (manufactured by Nippi Co., Ltd.), using StemFit AK03 or AK03N medium (manufactured by Ajinomoto Co., Ltd.) by a known feeder-free method (Nakagawa M, Taniguchi Y, Senda S, Takizawa N, Ichisaka T, Asano K, Morizane A, Doi D, Takahashi J, Nishizawa M, Yoshida Y, Toyoda T, Osafun...
Embodiment 2
[0195] [Example 2] Suicide gene cell therapy for glioma stem cells
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[0197] Human glioma stem cells were transplanted into the right striatum (2 mm to the right of bregma and 3 mm from the brain surface) of T cell-deficient mice (female BALB / c nude mice, 20 g, 6 weeks) after general anesthesia Strain hG008-ffLuc 5×10 5 cells / 2μl, transplanted yCD-UPRT expressing TSCs (using CRISPR / Cas9 to introduce yCD-UPRT into iPS cells) 1×10 at the same site after 45 days 6 each / 2μl. Seven days later, after the tumor was confirmed by the IVIS in vivo imaging system, the treatment group was intraperitoneally administered 5-FC (10 mg / ml) with 500 μl once / day for 2 weeks (TSC(+) / 5-FC(+) 5mg / day, n=8). The control group 1 was similarly administered with 500 µl of 1M PBS for 2 weeks (control group 1: TSC(+) / 5-FC(-), n=7). In addition, a group transplanted with only hG008-ffLuc was also prepared as another control group, and 5-FC (10 mg / ml) was intraperitoneally administered with ...
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