Optimization of in vitro model for analysis of tumor cell migration dynamics

Abstract

Migration capacity is an important feature of tumor cells. There are several approaches to analyze the dynamics of cancer cell migration in vitro. The model of initiation of cell migration from 3D multicellular spheroids onto growth surface is one of the closest to the in vivo conditions. Aim. Optimization of the model to study tumor cell mobility for several days. Methods. 2D and 3D MCF-7 cell culture, immunofluorescence analysis and image analysis using the Fiji computer software. Results. Unification of spheroid size allowed avoiding a significant data deviation. The obtained spheroids spread completely for three days. The highest migration ratio was observed on the second day. The proliferation level was similar during each day of the three-day experiment; it did not exceed 3 %. The validity of the model was tested after migration inhibition by a mTOR signaling inhibitor rapamycin. Additionally, this model was successfully applied to immunofluorescence study of p85S6K1 subcellular localization in moving MCF-7 cells. Conclusions. Double filtration of multicellular spheroids allowed unification of their size; this promotes an adequate interpretation of the migration assay. This model allows to study tumor cell migration dynamics and can be further used for development of anticancer drugs.

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