title: Vitreous cryopreservation of human preantral follicles encapsulated in alginate beads with mini mesh cups.
abstract: To completely avoid ice crystal formation and thus get a higher survival rate, vitrification methods have been commonly used for cryopreservation of oocytes and embryos. However, currently used vitrification methods for oocytes and embryos are not suitable for the cryopreservation of preantral follicles (PFs). In the present study, stainless steel mesh was fabricated into mini mesh cups to vitrify isolated PFs. Moreover, isolated follicles were encapsulated and then subjected to vitreous cryopreservation to facilitate in vitro culture/maturation of follicles after warming. The results showed that the percentages of viable follicles did not differ significantly between the vitrification group and fresh group soon after warming (81.25% vs. 85.29%, P>0.05) and after a 7-day culture period (77.78% vs. 83.33%, P>0.05). No difference in mean follicular diameter was observed between cryopreserved and fresh follicles when cultured in vitro. Transmission electron microscopic analysis revealed that vitreous cryopreservation could maintain the ultrastructure of follicles in alginate beads. In conclusion, the present vitrification method could efficiently cryopreserve isolated human ovarian follicles encapsulated by calcium alginate, which could be put into immediate use (in vitro culture/ maturation) after warming. However, more follicles and some detailed biochemical analyses are required to further investigate the effects of vitrification on the long-term growth of human encapsulated PFs.
authors.affiliation: Reproductive Medicine Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, PR China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Bian
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Chenhui
authors.initials: C
authors.lastname: Ding
authors.forename: Weijie
authors.initials: W
authors.lastname: Xin
authors.forename: Bo
authors.initials: B
authors.lastname: Zhu
authors.forename: Canquan
authors.initials: C
authors.lastname: Zhou
mesh_terms: D000328:Adult; D000464:Alginates / Q000737:chemistry*; D018929:Cell Culture Techniques; D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods*; D005260:Female; D005452:Fluoresceins / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry; D006107:Granulosa Cells / Q000166:cytology; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D009865:Oocytes / Q000166:cytology; D006080:Ovarian Follicle / Q000166:cytology* / Q000648:ultrastructure; D013193:Stainless Steel; D013936:Thymidine / Q000737:chemistry; D058989:Vitrification; D055815:Young Adult
chemical_list: D000464:Alginates; D005452:Fluoresceins; D006603:Hexuronic Acids; D013193:Stainless Steel; D020723:Glucuronic Acid; C007740:fluorexon; D013936:Thymidine