Monitoring the migration of bone marrow derived mesenchymal stem cells to intracranial glioma by sodium iodide sympoter

Abstract

Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene, and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma. Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro, then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP. pLVX-CMV-0-EGFP was constructed as control. BMSCs were isolated, cultured, and transfected by lentivirus. The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected. The expression of NIS gene was examined by Western blot. Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4. The nude mice intracranial glioma models were established. MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein. Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged. BMSCs were successfully isolated and cultured. Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection. The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP, but not in BMSCs-EGFP. BMSCs-NIS-EGFP showed significantly more uptake of 125I(nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4. The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection. Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma. It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma. Key words: Glioma; Bone marrow; Stem cells; Sodium/iodide symporter; Tomography, emission-computed, single-photon; Mice, nude