Abstract
Human embryonic kidney 293T (HEK293T) cells are used in various biological experiments and researches. In this study, we investigated the effect of cell culture environments on morphological and functional properties of HEK293T cells. We used several kinds of dishes made of polystyrene or glass for cell culture, including three types of polystyrene dishes provided from different manufacturers for suspension and adherent cell culture. In addition, we also investigated the effect of culturing on gelatin-coated surfaces on the cell morphology. We found that HEK293T cells aggregated and formed into three-dimensional (3-D) multicellular spheroids (MCS) when non-coated polystyrene dishes were used for suspension culture. In particular, the non-coated polystyrene dish from Sumitomo bakelite is the most remarkable characteristic for 3-D MCS among the polystyrene dishes. On the other hand, HEK293T cells hardly aggregated and formed 3-D MCS on gelatin-coated polystyrene dishes for suspension culture. HEK293T cells adhered on the non- or gelatin-coated polystyrene dish for adherent culture, but they did not form 3-D MCS. HEK293T cells also adhered to non- or gelatin-coated glass dishes and did not form 3-D MCS in serum-free medium. These results suggest that HEK293T cells cultured on non-coated polystyrene dish may be useful for the tool to analyze the characteristics of 3D-MCS.
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Acknowledgements
We would like to thank our corroborators in our laboratory.
Funding
This research was funded by the Grant-in-Aid for Scientific Research of Seikei University (Grant No. 2018) and Scientific Research from the Japan Society for the Promotion of Science (18K11001 to KI).
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KI, KO, KH, YS, TN and HH conceived and designed the experiments, and contributed to manuscript preparation. KI, KO, KH, YS, and KS performed the experiments. All authors read and approved the final manuscript.
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10616_2019_363_MOESM1_ESM.pptx
Supplementary material 1 DLD-1 and SUIT-2 cells cultured on polystyrene dishes for suspension culture. DLD-1 and SUIT-2 cells were cultured on non-coated or gelatin-coated suspension culture dishes (dish-S) for 2 days. (PPTX 3389 kb)
10616_2019_363_MOESM2_ESM.pptx
Supplementary material 2 HEK293T cells cultured on low cell binding surface plates. HEK293T cells were cultured on Low Cell Binding Surface for 2 days (PPTX 1614 kb)
10616_2019_363_MOESM3_ESM.pptx
Supplementary material 3 HEK293T cells cultured on non-coated suspension culture dish. (A) HEK293T cells were cultured on non-coated suspension culture dish (dish-S) for 7 days. The cells were observed using phase contrast microscopy. (B) HEK293T cells were cultured on a non-coated suspension culture dish (dish-S) or an adherent culture dish (dish-TA) for 7 days. The cells were treated with Accutase for 1 h..Cell death was measured by staining with Hoechst 33342 and propidium iodide. (C) HEK293T cells were cultured on a non-coated suspension culture dish (dish-S) or an adherent culture dish (dish-TA) for 7 days. The cells were directly stained with Hoechst 33342 and propidium iodide and were observed under a microscope (PPTX 61500 kb)
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Iuchi, K., Oya, K., Hosoya, K. et al. Different morphologies of human embryonic kidney 293T cells in various types of culture dishes. Cytotechnology 72, 131–140 (2020). https://doi.org/10.1007/s10616-019-00363-w
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DOI: https://doi.org/10.1007/s10616-019-00363-w