Extended Data Fig. 2: Comparison of in vivo and in vitro grown oocytes.
From: Rejuvenation of aged oocyte through exposure to young follicular microenvironment
a, Diameter of oocytes grown in vivo or in vitro. n = 109 (in vivo), 90 (in vitro). b, Quantification of oocyte maturation rate. Sample sizes: n = 126 (in vivo) and n = 98 (in vitro) oocytes, with 4 biological replicates in each group. Data are shown as mean ± SD. c, Analysis of embryo development potential. n = 83 (in vivo) and 75 (in vitro). d-f, Transcriptome analysis of oocytes grown in vitro and in vivo. Volcano plot (d) of DEGs (p.adjust < 0.05 and log2 fold change > 0.5 or < −0.5) between in vitro and in vivo oocytes. Two-sided Wald-test adjusted with Benjamini-Hochberg method. Correlation heatmap (e) with hierarchical clustering to show the sample-to-sample distances. PCA analysis (f) of the normalized gene expression data. Ellipses fit a multivariate t-distribution at confidence level of 0.8. n = 8 in vivo and 8 in vitro. g, Dot plots illustrating follicle size changes over time during 3D ex vivo culture. Color bar and circle size represent follicle size. 2-month-old (young, n = 18) and 14-month-old (aged, n = 18) mice were used. h,i, Representative images (h) of 3D ex vivo cultured young and aged follicles. Follicles were considered atretic if there was disruption of contact between the oocyte (red asterisk) and GCs, leading to the release of oocytes from the follicles (bottom left), or if the follicles contained apoptotic or dead oocytes (bottom right). Antrum is indicated by the white arrowhead. Scale bar, 100 μm. Atresia rate was quantified (i) in young (2-3 months) and aged (14-15 months) follicles after 3D ex vivo culture. The median is represented by the center line, with individual dots representing biological replicates for each group. Sample sizes: n = 166 (young), 199 (aged) follicles, with 5 biological replicates in each group. 2-3 month-old mice were used in (a-f). Box plots inside the violins in (a) show mean (black circle), quartiles (box limits), and 1.5× interquartile range (whiskers). Two-tailed unpaired t-tests for (a, b, i). Two-tailed Fisher’s exact test for (c). P value: **P < 0.01, ns, not significant (P > 0.05). Exact P values are in the Source Data. All data are from at least three independent experiments.
