Quantitative analysis of cell tracing by in vivo imaging system

Abstract

In vivo imaging system (IVIS) is a new and rapidly expanding technology, which has a wide range of applications in life science such as cell tracing. By counting the number of photons emitted from a specimen, IVIS can quantify biological events such as tumor growth. We used B16F10-luc-G5 tumor cells and 20 Babl/C mice injected subcutaneously with B16F10-luc-G5 tumor cells (1×10⁶ in 100 μL) to develop a method to quantitatively analyze cells traced by IVIS in vitro and in vivo, respectively. The results showed a strong correlation between the number of tumor cells and the intensity of bioluminescence signal (R²=0.99) under different exposure conditions in in vitro assay. The results derived from the in vivo experiments showed that tumor luminescence was observed in all mice by IVIS at all days, and there was significant difference (P<0.01) between every two days from day 3 to day 14. Moreover, tumor dynamic morphology could be monitored by IVIS when it was invisible. There was a strong correlation between tumor volume and bioluminescence signal (R²=0.97) by IVIS. In summary, we demonstrated a way to accurately carry out the quantitative analysis of cells using IVIS both in vitro and in vivo. The data indicate that IVIS can be used as an effective and quantitative method for cell tracing both in vitro and in vivo.

Keywords

in vivo imaging system / cell tracing / quantitative analysis

References

[1] GreerL.F., SzalayA.A.. Imaging of light emission from the expression of luciferases in living cells and organisms: a review. Luminescence, 2002, 17(1): 43-74

[2] MassoudT.F., GambhirS.S.. Molecular imaging in living subjects: seeing fundamental biological processes in a new light. Genes Dev, 2003, 17(5): 545-580

[3] LarmonierN., JanikashviliN., LaCasseC.J., et al.. Imatinib mesylate inhibits CD4+ CD25+ regulatory T cell activity and enhances active immunotherapy against BCR-ABL-tumors. J Immunol, 2008, 181(10): 6955-6963

[4] HanZ., FuA., WangH., et al.. Noninvasive assessment of cancer response to therapy. Nat Med, 2008, 14(3): 343-349

[5] XueW., ZenderL., MiethingC., et al.. Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature, 2007, 445(7128): 656-660

[6] TammelaT., SaaristoA., HolopainenT., et al.. Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation. Nat Med, 2007, 13(12): 1458-1466

[7] StephanM.T., PonomarevV., BrentjensR.J., et al.. T cell-encoded CD80 and 4-1BBL induce auto- and transcostimulation, resulting in potent tumor rejection. Nat Med, 2007, 13(12): 1440-1449

[8] DentinR., LiuY., KooS.H., et al.. Insulin modulates gluconeogenesis by inhibition of the coactivator TORC2. Nature, 2007, 449(7160): 366-369

[9] ZhangL., LeeK.C., BhojaniM.S., et al.. Molecular imaging of Akt kinase activity. Nat Med, 2007, 13(9): 1114-1119

[10] DierksC., GrbicJ., ZirlikK., et al.. Essential role of stromally induced hedgehog signaling in B-cell malignancies. Nat Med, 2007, 13(8): 944-951

[11] BarberiT., BradburyM., DincerZ., et al.. Derivation of engraftable skeletal myoblasts from human embryonic stem cells. Nat Med, 2007, 13(5): 642-648

[12] HajitouA., RangelR., TrepelM., et al.. Design and construction of targeted AAVP vectors for mammalian cell transduction. Nat Prot, 2007, 2(3): 523-531

[13] MurphyG.J., MostoslavskyG., KottonD.N., et al.. Exogenous control of mammalian gene expression via modulation of translational termination. Nat Med, 2006, 12(9): 1093-1099

[14] ZakrzewskiJ.L., KochmanA.A., LuS.X., et al.. Adoptive transfer of T-cell precursors enhances T-cell reconstitution after allogeneic hematopoietic stem cell transplantation. Nat Med, 2006, 12(9): 1039-1047

[15] NegrinR.S., ContagC.H.. In vivo imaging using bioluminescence: a tool for probing graft-versus-host disease. Nat Rev, 2006, 6(6): 484-490

[16] Franke-FayardB., WatersA.P., JanseC.J.. Real-time in vivo imaging of transgenic bioluminescent blood stages of rodent malaria parasites in mice. Nat Prot, 2006, 1(1): 476-485

[17] WehrmanT.S., von DegenfeldG., KrutzikP.O., et al.. Luminescent imaging of β-galactosidase activity in living subjects using sequential reporterenzymeluminescence. Nat Methods, 2006, 3(4): 295-301

[18] ThorneS.H., NegrinR.S., ContagC.H.. Synergistic antitumor effects of immune cell-viral biotherapy. Science, 2006, 311: 1780-1784

[19] MinnA.J., GuptaG.P., SiegelP.M., et al.. Genes that mediate breast cancer metastasis to lung. Nature, 2005, 436(28): 518-524

[20] MorizonoK., XieY., RingpisG.E., et al.. Lentiviral retargeting to P-glycoprotein on metastatic melanoma through intravenous injection. Nat Med, 2005, 11(3): 346-352

[21] KimJ.H., KimB., CaiL., et al.. Transcriptional regulation of a metastasis suppressor gene by Tip60 and β-catenin complexes. Nature, 2005, 434(7035): 921-926

[22] HoebeK., GeorgelP., RutschmannS., et al.. CD36 is a sensor of diacylglyerides. Nature, 2005, 433(7025): 523-527

[23] BinsA.D., JorritsmaA., WolkersM.C., et al.. A rapid and potent DNA vaccination strategy defined by in vivo monitoring of antigen expression. Nat Med, 2005, 11(8): 899

[24] GrossS., Piwnica-WormsD.. Real-time imaging of ligand-induced IKK activation in intact cells and in living mice. Nat Methods, 2005, 2(8): 607-614

[25] LevenbergS., RouwkemaJ., MacdonaldM., et al.. Engineering vascularized skeletal muscle tissue. Nat Biotechnol, 2005, 23(7): 879-884

[26] GreerL.F., SzalayA.A.. Imaging of light emission from the expression of luciferases in living cells and organisms: a review. Luminescence, 2002, 17(1): 43-74