Extended Data Fig. 3: Growth and maturation of oocytes from RCFs in 3D ex vivo culture.
From: Rejuvenation of aged oocyte through exposure to young follicular microenvironment
a, Procedure for generating reconstituted chimeric follicles. Red arrow points to the oocyte used for transplantation. Red asterisk indicates the oocyte within the r-follicle that will be replaced. Refer to Supplementary Video 1 and Methods for further details. b, To distinguish between the donor oocyte and the r-follicle, we employed oocytes from mTmG transgenic mice exhibiting membrane-localized tdTomato (pseudo-colored yellow). In contrast, the r-follicles were sourced from non-fluorescent wild-type mice. The mTmG oocytes served as donors as referenced in Fig. 2g and Extended Data Fig. 3c–e. c, RCF size increased during 3D ex vivo culture. Oocytes from transgenic mTmG mice and follicular somatic cells from wild-type mice, as shown in (b). Scale bars, 50 μm. d, Cumulus-oocyte complexes (COCs) isolated from antral RCFs were induced for oocyte maturation with hCG for 16 hours in vitro. Note that cumulus cells surrounding the oocytes (from mTmG mice) expanded, and oocytes resumed meiosis, extruded the PB1 as shown in (e). Scale bars, 200 μm. e, Representative image of mature eggs derived from RCFs as shown in (c and d). The cumulus cells were removed after maturation to visualize mature eggs with the first polar body (PB1, arrows). Scale bars, 40 μm. All images are representative of at least three independent experiments.
