Fig. 3: Effect of MIH and LIH on PAFs during warming.

a Differential interference contrast (DIC) and fluorescence images of PAFs after warming under the following different conditions: W/ Encap or W/O Encap, W/ LIH, W/ MIH or W/ MIH and LIH during warming. b Viability of the PAFs after vitrification with different CPAs (0 M, green; 1 M, light green; 1.5 M, orange; 2 M, purple bars). P_1–1.5 = 0.5978, P_1–2 = 0.4151, P_1.5–2 = 0.9785. Each experiment was repeated three times with similar results, and representative images are shown. c Viability of the PAFs encapsulated in different concentrations of alginate hydrogel (0%, green; 1%, rust; 1.5%, pink; and 2%, emerald green bars. w/v) after vitrification with different 1.5 M CPA. P_1–1.5 = 0.9553, P_1–2 = 0.7018, P_1.5–2 = 0.9894. d Viability of PAFs after warming under different conditions (Fresh: green; W/O Encap, MIH and LIH: light green; W/ Encap: orange; W/ Encap and MIH: pink; W/ Encap and LIH: emerald green; W/ Encap, MIH and LIH: blue bars). P_{Fresh- W/ MIH + LIH} = 0.1740, P_{W/O Enap-W/ Enap, W/O MIH or LIH} = 0.0265, P_{W/ Enap, W/O MIH or LIH-W/ MIH} = 0.0306, P_{W/ Enap, W/O MIH or LIH-W/ LIH} = 0.0221, P_{W/ MIH-W/ MIH +  LIH} = 0.0137, P_{W/ LIH-W/ MIH + LIH} = 0.0045. 1 M: 0.5 M EG + 0.5 M PROH + 1 M trehalose; 1.5 M: 0.75 M EG + 0.75 M PROH + 1 M trehalose; 2 M: 1 M EG + 1 M PROH + 1 M trehalose. W/O Encap: without encapsulation; W/ Encap: with encapsulation; W/ MIH: with magnetic induction heating; W/O MIH: without magnetic induction heating; W/ LIH: with laser-induced heating; W/O LIH: without laser-induced heating. W/ MIH + LIH: with magnetic induction heating and laser-induced heating. One-way analysis of variance (ANOVA) and Tukey’s post hoc were used for statistical analysis. ns: p > 0.05; *p ≤ 0.05; **p ≤ 0.01. n = 18–23 for four replicates. n: number of follicles used in each experiment. Data are presented as the mean ± SD (b, c, d).