title: Slow-controlled freezing versus speed-cooling for cryopreservation of whole guinea pig ovaries.
abstract: The objective of this study was to evaluate the feasibility of whole-ovary perfusion, and to compare the effects of speed-cooling and slow-controlled freezing of whole guinea pig ovaries. Slow-freezing and speed-cooling procedures were performed after perfusion of guinea pig ovaries with cryoprotectants. Ink perfused via the vascular pedicles was present in the microvessels around various follicles at various stages of development in the cortical and medullar regions, thereby confirming that perfusion was effective. Vascular damage was essentially confined to the cannulated artery. Based on histological examination, there were (mean ± SEM) 93.1 ± 4.2, 79.0 ± 2.0, and 54.7 ± 8.5% healthy follicles in the fresh, slow-freezing and speed-cooling groups, respectively (each group differed from the other two, P < 0.05). Trypan blue staining of isolated follicles confirmed that cellular damage was greater following speed-cooling than slow-freezing (58.6 vs 29.2%, P < 0.05). Based on a TUNEL assay, speed-cooling caused more apoptotic granulosa and theca cells in antral follicles than slow-freezing. In conclusion, the present study provided evidence that guinea pig whole ovaries could be perfused with cryoprotectant and cryopreserved in vitro. Furthermore, the slow-freezing protocol resulted in less cellular damage in thawed tissues than speed-cooling.
authors.affiliation: Centre for Reproductive Medicine, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Xu
authors.forename: Xiaojiao
authors.initials: X
authors.lastname: Wang
authors.forename: Yonggen
authors.initials: Y
authors.lastname: Wu
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Meng
authors.forename: Fangfang
authors.initials: F
authors.lastname: Wu
authors.forename: Na
authors.initials: N
authors.lastname: Zhou
authors.forename: Wenwei
authors.initials: W
authors.lastname: Chen
authors.forename: Bilu
authors.initials: B
authors.lastname: Ye
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D006168:Guinea Pigs; D010053:Ovary / Q000473:pathology / Q000502:physiology*; D014021:Tissue Preservation / Q000379:methods*