Preparation and Testing of Cells Expressing Fluorescent Proteins for Intravital Imaging of Tumor Microenvironment

K. K. Sukhinich; A. V. Makarov; V. A. Naumenko; M. A. Abakumov; A. G. Majouga; S. S. Vodopyanov; M. A. Kunin; A. S. Garanina; N. F. Grinenko; K. Yu. Vlasova; P. A. Mel’nikov; V. P. Chekhonin

doi:10.1007/s10517-019-04475-3

Preparation and Testing of Cells Expressing Fluorescent Proteins for Intravital Imaging of Tumor Microenvironment

Authors: Sukhinich K.K.¹, Makarov A.V.², Naumenko V.A.³, Abakumov M.A.³^,4, Majouga A.G.³^,5^,6, Vodopyanov S.S.³, Kunin M.A.⁵, Garanina A.S.³, Grinenko N.F.², Vlasova K.Y.⁵, Mel’nikov P.A.², Chekhonin V.P.²^,4
Affiliations:
1. N. K. Kol’tsov Institute of Developmental Biology, Russian Academy of Sciences
2. V. P. Serbsky Federal Medical Research Center for Psychiatry and Narcology, Ministry of Health of the Russian Federation
3. Laboratory of Biomedical Nanomaterials, National University of Science and Technology (MISIS)
4. N. I. Pirogov Russian National Research Medical University, Ministry of Health of the Russian Federation
5. M. V. Lomonosov Moscow State University
6. D. I. Mendeleev University of Chemical Technology
Issue: Vol 167, No 1 (2019)
Pages: 123-130
Section: Translated from Kletochnye Tekhnologii v Biologii i Meditsine (Cell Technologies in Biology and Medicine)
URL: https://journals.rcsi.science/0007-4888/article/view/241509
DOI: https://doi.org/10.1007/s10517-019-04475-3
ID: 241509

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Abstract

Intravital microscopy is widely used for in vivo studies of the mechanisms of carcinogenesis and response to antitumor therapy. For visualization of tumor cells in vivo, cell lines expressing fluorescent proteins are needed. Expression of exogenous proteins can affect cell growth rate and their tumorigenic potential. Therefore, comprehensive analysis of the morphofunctional properties of transduced cells is required for creating appropriate models of tumor microenvironment. In the present study, six lines of mouse tumor cells expressing green and red fluorescent proteins were derived. Analysis of cells morphology, growth kinetics, and response to chemotherapy in vitro revealed no significant differences between wild-type and transduced cell lines. Introduction of fluorescent proteins into the genome of 4T1 (murine breast cancer) and B16-F10 (murine melanoma) cells did not affect tumor growth rate after subcutaneous implantation to mice, while both CT26-GFP and CT26-RFP cells (murine colon cancer) were rejected starting from day 8 after implantation. Elucidation of the mechanisms underlying CT26-GFP/RFP rejection is required to modify transduction technique for creating the models of tumor microenvironment accessible for in vivo visualization. Transduced 4T1 and B16-F10 cell lines can be used for intravital microscopic imaging of tumor cells, neoplastic vasculature, and leukocyte subpopulations.

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Preparation and Testing of Cells Expressing Fluorescent Proteins for Intravital Imaging of Tumor Microenvironment

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Abstract

About the authors

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