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. 2020 Aug 12;10:13605. doi: 10.1038/s41598-020-70707-6

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of nano-warming on cryopreservation of hiPSC aggregates. (a) Schematic illustration for cryopreservation of iPSC aggregates. hiPSCs were applied to a 30-mL stirred bioreactor at 5 × 10⁵ cells/mL. The cells were cultured for 2 days to form hiPSC aggregates. For cryopreservation of iPSC aggregates, StemCell Keep containing magnetite nanoparticles (5 mg/mL) was added to the cell aggregates at 1 × 10⁶ cells/mL in glass vials, and the vials were directly immersed in liquid nitrogen. (b) Effects of nano-warming on the viability of hiPSC aggregates. After freezing and thawing, cell viability was assayed using a cell viability imaging kit based on Hoechst 33342 for live cells and SYTOX green nucleic acid stain for dead cells. White columns, convective warming (water bath at 37 °C) of StemCell Keep; grey columns, convective warming (water bath at 37 °C) of magnetite-loaded StemCell keep; black columns, nano-warming of magnetite-loaded StemCell keep. Data are expressed as the mean ± SD (n = 3). *P < 0.05. N.S not significant. (c) Florescence images of hiPSC aggregates in non-frozen control (top) and nano-warming (bottom) groups, showing nuclei (DAPI) and Oct3/4. Scale bars, 50 µm.