In Vivo Bioluminescence Imaging of Tumor Cells Using Optimized Firefly Luciferase luc2

The present study was aimed to establish a tumor cell line stably expressing luciferase luc2, and to develop the technique to observe primary tumor nodes and metastases using in vivo bioluminescence imaging.

Materials and Methods. In this research we used pLuc2-N plasmid, lentiviral vector pLVT-1, Colo 26 cell line and BALB/c mice to generate new bioluminescent tumor model. Bioluminescence imaging in vitro и in vivo was carried out on IVIS-Spectrum system (Caliper Life Sciences, USA). Primary tumor model was created by subcutaneous injection of 500 000 Colo 26-luc2 cells. Model of metastases was generated by i.v. injection of 75 000 Colo 26-luc2 cells. Histological analysis was performed to verify the results of the imaging.

Results. We created the lentiviral vector containing luc2 gene using molecular cloning. Then Colo 26-luc2 tumor cell line was generated. We assessed the sensitivity of luc2-based bioluminescence imaging. The intensity of bioluminescent signal in vitro averaged about 5000 photon/s per cell, in vivo — 250 photon/sec per cell. In vivo monitoring of Colo 26-luc2 primary tumor and metastases was demonstrated. The results of bioluminescence imaging correlated with histological analysis data.

Conclusion. The present work shows the possibility of bioluminescent system based on optimized luciferase luc2 for in vivo noninvasive high-sensitive whole-body imaging of tumors.

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Klementyeva N.V., Shirmanova M.V., Serebrovskaya Е.О., Fradkov А.F., Meleshina А.V., Snopova L.B., Prodanets N.N., Lukyanov S.А., Zagaynova Е.V. In Vivo Bioluminescence Imaging of Tumor Cells Using Optimized Firefly Luciferase luc2. Sovremennye tehnologii v medicine 2013; 5(3): 6