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. 2023 Jan 9;15(2):2630–2638. doi: 10.1021/acsami.2c18288

Figure 2.

Figure 2

Post-thaw (24 h) recovery of cryopreserved HepG2 spheroids. Percentage viability of spheroids (A) frozen directly in agar molds and stored at −80 °C for 24 h; (B) frozen suspended in cryovials and stored at −80 °C for 24 h; and (C) frozen in cryovials and stored above liquid nitrogen for 3 days. Data are presented as mean % viability relative to pre-frozen spheroids ± SEM from five independent repeats, determined by the WST-1 assay. (D) Confocal microscopy of spheroids before and after thawing in DMSO (10%) with and without supplementation with 20 mg mL–1 polyampholyte. Living cells are labeled green (calcein-AM, intact membrane), dead cells are labeled red (EthD-III, damaged membrane), and the nuclei of cells with intact membranes were stained with Hoechst 33342 solution (20 mM). Scale bar = 100 μm.