Author contributions: J.M-Q: conception and design, data collection and assembly, data analysis and interpretation, and manuscript writing; D.B. and D.H.: data collection and assembly; P.H. and U.M.: provision of study material and data collection and analysis; K.S.: conception and design, data analysis and interpretation, financial support, and manuscript writing.
Translational and Clinical Research
Therapeutic Efficacy and Fate of Bimodal Engineered Stem Cells in Malignant Brain Tumors†
Version of Record online: 23 AUG 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 8, pages 1706–1714, August 2013
How to Cite
Martinez-Quintanilla, J., Bhere, D., Heidari, P., He, D., Mahmood, U. and Shah, K. (2013), Therapeutic Efficacy and Fate of Bimodal Engineered Stem Cells in Malignant Brain Tumors. STEM CELLS, 31: 1706–1714. doi: 10.1002/stem.1355
- Issue online: 23 AUG 2013
- Version of Record online: 23 AUG 2013
- Accepted manuscript online: 6 FEB 2013 11:29PM EST
- Manuscript Accepted: 31 DEC 2012
- Manuscript Received: 31 MAY 2012
Additional Supporting Information may be found in the online version of this article.
|STEM_1355_sm_SuppFig1.tif||1950K||Supplemental Fig.1. Expression of bimodal imaging transgenes using engineered lentiviral and retroviral vectors. (A) A self-inactivating lentiviral and retroviral transfer vectors, both bearing an IRES-GFP element, were used to construct the following vectors: LV-TK, LVTR , RV-GFl, RV-TK and RV-TR under the CMV promoter. (B) Serial dilutions of mMSC-TK cells expressing GFP-Fluc (mMSC-TK-GFl) were plated and 24hrs later Fluc signal intensity was determined. Plot shows direct correlation between mMSC-TK-GFl cell number and Fluc signal intensity. Abbreviations: CMV, cytomegalovirus promoter; Fluc, firefly luciferase; IRES, Internal ribosomal entry site; GFl, green fluorescent protein and firefly luciferase fusion; TK, Thymidine Kinase; TRAIL, tumor necrosis factor apoptosis-inducing ligand.|
|STEM_1355_sm_SuppFig2.tif||1745K||Supplemental Fig.2. hMSC-TR-TK have antitumor effect and can be eliminated post tumor treatment in vitro. U87-FmC cells were co-cultured with different proportions of hMSCTR (grey columns) or hMSC-TR-TK (black columns) and two days later, GCV (10μg/mL) was added to the cultures. (A) Plot shows the activation of Caspase-3/7 in the co-cultures with U87- FmC and engineered hMSC at day one. (B) Plot shows U87-FmC viability measured by Fluc signal in co-culture conditions at day three. (C-G) U87-FmC were co-cultured with engineered hMSC-TR-GFl or hMSC-TR-TK-GFl and two days later GCV (10μg/mL) was added to the cultures. Photomicrographs (original magnification, x4) show hMSC GFP+ cells at day three post GCV treatment (C-F). (G) Plot shows % of viability of modified hMSC-Fluc cells measured by Fluc signal at day three. Bars, +SD. In all panels, *, p < .05 versus controls. Abbreviations: hMSC, human mesenchymal stem cells; GCV, ganciclovir; TK, Thymidine Kinase; TR, tumor necrosis factor apoptosis-inducing ligand; Fluc, firefly luciferase; GFl, green fluorescent protein and Fluc fusion.|
|STEM_1355_sm_SuppFig3.tif||407K||Supplemental Fig.3. Intratumorally implantation and fate of mMSC-TR-TK. Gli36vIII-FmC bearing mice were intratumorally implanted with mMSC-TR-TK. Photomicrographs of mice brain sections showing presence of therapeutic mMSC-TR-TK (green) in GBM tumors (mcherry) after 4 days post mMSC implantation (A). Photomicrographs of mice brain sections showing CD31 staining (red) and mMSC-TR-TK (green) at day 32 post-implantation (B). (Original magnification, x10 left panel and x20 right panel).|
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