Mouse ovarian follicle cryopreservation using vitrification or slow programmed cooling: Assessment of in vitro development, maturation, ultra-structure and meiotic spindle organization

Abstract

Isolated follicles were more vulnerable to cryodamage after slow freezing as compared to vitrification.

A simple and efficient method for preparation of isolated ovarian follicles for transmission electron microscopy

“…The polyester membranes, transfixed to the transwell inserts, are available in 3 pore sizes; 0.4 µm, 0.8 µm and 3 µm. Transwell inserts have been used for in vitro culture and maturation of ovarian follicles [24][25][26]. The polyester membrane is fixed to the bottom of the insert, onto which ovarian follicles are cultured.…”

“…In contrast, the polyester membrane in the transwell insert is translucent, allowing excellent cell visualization under phase contrast microscopy without the need for staining. The transwell insert with polyester membrane is a well documented system for in vitro culture and maturation of ovarian follicles [24][25][26]. The major limitation with this technique is that the original three-dimensional structure of the ovarian follicles is lost by attachment to the transwell membrane.…”

Three dimensional culture of fresh and vitrified mouse pre-antral follicles in a hyaluronan-based hydrogel: a preliminary investigation of a novel biomaterial for in vitro follicle maturation

“…The protocol for vitrification and warming of isolated pre-antral follicles has been previously described in detail [19]. The basal medium for all vitrification solutions was L-15 with 20% synthetic serum substitute (SSS; Irvine Scientific; Irvine, CA).…”

Total Oxidative status of Mouse Vitrified Pre-Antral Follicles with Pre-Treatment of Alpha Lipoic Acid

“…The survival rate of the pre-antral follicles was assessed microscopically based on the morphology of the pre-antral follicle every other day under an inverted microscope during culturing period. Then, they were compared at the end of the study as described previously [16]. Briefly, a follicle was considered normal when it possessed a centrally located spherical and also homogeneous oocyte surrounded by complete and compact layers of granulosa cells without noticeable damage to the basement membrane.…”