Intravital optical bioimaging of ovarian cancer using a luminescent cell line

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Abstract

The method of intravital bioimaging based on luminescence occupies an important place in the development and testing of antitumor drugs on model animals and is an essential part of preclinical studies. Bioimaging based on luminescent systems, compared with fluorescent bioimaging, provides a high signal-to-noise ratio, which justifies the development of cell lines, that stably express luciferase genes for subsequent use in model animals. The work describes the creation of a stable cell line SKOV3.ip1-NanoLuc constitutively expressing the NanoLuc luciferase gene. The developed cell line was shown to be effective for intravital luminescence bioimaging of immunodeficient animals with deep-seated intraperitoneal tumors, which can be considered as a model of late-stage ovarian cancer.

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About the authors

E. I. Shramova

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of science

Author for correspondence.
Email: shramova.e.i@gmail.com
Russian Federation, Moscow

G. M. Proshkina

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of science

Email: shramova.e.i@gmail.com
Russian Federation, Moscow

S. M. Deyev

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of science; National Research Center “Kurchatov Institute”; Kazan Federal University

Email: shramova.e.i@gmail.com

Academician of the RAS, “Biomarker” Research Laboratory, Institute of Fundamental Medicine and Biology

Russian Federation, Moscow; Moscow; Kazan

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Supplementary files

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2. Fig. 1. Characterization of the stable cell line SKOV3.ip1/NanoLuc. A – luminescence spectrum of SKOV3.ip1/NanoLuc cells in the presence of furimazine; B – graph of the photon flux value versus the number of cells, obtained using the IVIS Spectrum CT intravital bioimaging system. The inset shows a photograph of luminescent signals of different numbers of cells, obtained using the IVIS system; C – results of cytofluorometric analysis of parental SKOV3.ip1 cells (left histogram) and SKOV3.ip1/NanoLuc cells (right) treated with FITC-labeled trastuzumab. In both histograms, the red curves correspond to the signal from cells not treated with the antibody (autofluorescence), the blue curves correspond to the signal from cells treated with trastuzumab. The mean fluorescence intensity (MFI) values ​​are shown for each curve. The FITC (fluorescein isothiocyanate) channel corresponds to the FITC detection parameters λex = 488 nm, λem = 530±30 nm; G is a superimposed confocal image of SKOV3.ip1/NanoLuc cells in the green and blue fluorescence channels after incubation with FITC-labeled trastuzumab. Nuclei are stained with Hoechst33342.

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3. Fig. 2. Monitoring the growth of intraperitoneal tumor nodules of SKOV3.ip1/NanoLuc cells in immunodeficient mice. A – fluorescent images of animals obtained using the IVIS Spectrum CT intravital bioimaging system on days 7, 14, 21, and 28 after intraperitoneal injection of cells. B – a diagram of tumor foci growth constructed based on the values ​​of fluorescent signal intensity in the abdominal cavity at the corresponding time point. The dotted line indicates the exponential approximation line of the data and the approximation reliability value (R2).

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