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. Author manuscript; available in PMC: 2010 Dec 1.
Published in final edited form as: Cancer Res. 2009 Nov 17;69(23):8932–8940. doi: 10.1158/0008-5472.CAN-08-3873

Figure 5.

Figure 5

Optimization of delivery of Delta-24-RGD to U87MG xenografts via hMSCs. A, An increased number of delivered hMSCs correlates with an increased number of engrafted hMSCs. Mice were implanted with 5 × 105 U87MG cells in the right frontal lobe and allowed to grow for 7 days. 1.0, 1.5, or 2 × 106 GFP-labled-hMSCs were injected into the carotid artery. Mice were sacrificed 4 days later. H&E scale bar 2 mm, hMSC-GFP scale bar 100 μm. B, (a) hMSCs were infected with Delta-24-RGD at 10, 50, or 100 MOI and allowed to incubate for 24, 48, or 72 hours. Cells were collected along with media and the titer of Delta-24-RGD was measured to determine the number of infectious units released. The highest number of infectious units was obtained using 100 MOI incubated for 48 hours. Experiment performed in triplicate. (b) hMSCs were plated in serum-free media and allowed to synchronize. At 0 hours, cells were infected with Delta-24-RGD at 0, 10, 50, or 100 MOI. At the indicated time points, cell cycle analysis was performed. Results are the percentage of cells in S-phase or pseudo-S-phase. All hMSCs infected with 50 or 100 MOI exhibited evidence of viral DNA replication by 48 hours. (c) hMSCs were infected with Delta-24-RGD at the indicated MOIs, then washed and plated in the upper wells of a Transwell plate (1×104cells/well). U87MG cells were plated in the bottom wells (3×104cells/well) and incubate for 7 days, after which U87MG cells were counted. Killing was seen with MOIs ≥ 10 and incubation times of ≥24hrs. C, Mice were implanted with U87MG cells and treated with hMSCs-Delta24 at the indicated MOIs and incubation times. Four days later, brains were analyzed by immunofluorescence for hexon protein expression (red cells). The 50MOI × 24h and 100MOI × 48h incubations produced equivalent infections. Scale bar 100μm. D, Mice were implanted with U87MG cells and injected IC with 1.5×106 hMSCs-Delta24 (50 MOI×24h incubation) 3 (upper) or 4 (lower) days later. On day 7 after delivery of hMSCs, xenografts were analyzed for hexon protein expression (green in top panel and red in bottom panel). H&E scale bar 1 mm, hexon scale bar 200 μm.