Comparison of different cryoprotectants in whole sheep ovary cryopreservation
Author(s): DU Tian-qi, CHI Ling-long, ZHAO Shu-qin, SHI Qing, QIN Rui-xi, SUN Yan-yan, LI Dong, DENG Xiao-hui, Infertility Center, Qilu Hospital of Shandong University, Shandong Qilu Stem Cells Co. Ltd, Zaozhuang Maternal and Child Care Service Hospital, Cryomedicine Laboratory, Qilu Hospital of Shandong University, Department of Pathology, Qilu Hospital of Shandong University
Pages: 415-421
Year: 2015 Issue: 3
Journal: Acta Anatomica Sinica
Keyword: Ovary; Cryopreservation; Trehalose; Glycerin; DMSO; TUNEL; Sheep;
Abstract: Objective To study the effects of freezing sheep intact ovary by perfusing different cryoprotectants with concentration gradient blended programmed organ cooling perfusor,and to obtain the best cryoprotectants combination for cryopreservation of the whole sheep ovary. Methods Twenty-eight ovaries collected from 6-8 months non-pregnant female sheeps were randomly distributed into fresh group( group A),trehalose group( group B),glycerin group( group C) and DMSO group( group D). The morphology,cell apoptosis( by HE staining and TUNEL assay) and mRNA transcript of Bcl-2 associated X protein and cold inducible RNA-binding protein( by real-time PCR) of thawed sheep ovaries were tested to established the criterion for appraising cryopreservation results. Results The percentage of normal follicles in group B was comparable with group A( P > 0. 05). The value in group C and group D were significant lower than that in group A( P < 0. 05). Quantitative assessment of stromal cell density indicated that the values in group D significantly lower than values in group A while group B and group C had values similar to those of group A. The TUNEL assay showed that the number of positive cells in group A were the lowest of all groups followed by group B,group C and group D( P < 0. 05).The level of BAX transcripts significantly increased in group C and group D( P < 0. 05). The level of CIRP transcripts increased highest in group B followed by group C and group D( P < 0. 05). Conclusion The whole ovary can be appropriately preserved after cryopreservation, and cryoprotectant combination of group B appears to be better for cryopreservation of whole sheep ovary in aspect of histology and anti-apoptosis.
Pages: 415-421
Year: 2015 Issue: 3
Journal: Acta Anatomica Sinica
Keyword: Ovary; Cryopreservation; Trehalose; Glycerin; DMSO; TUNEL; Sheep;
Abstract: Objective To study the effects of freezing sheep intact ovary by perfusing different cryoprotectants with concentration gradient blended programmed organ cooling perfusor,and to obtain the best cryoprotectants combination for cryopreservation of the whole sheep ovary. Methods Twenty-eight ovaries collected from 6-8 months non-pregnant female sheeps were randomly distributed into fresh group( group A),trehalose group( group B),glycerin group( group C) and DMSO group( group D). The morphology,cell apoptosis( by HE staining and TUNEL assay) and mRNA transcript of Bcl-2 associated X protein and cold inducible RNA-binding protein( by real-time PCR) of thawed sheep ovaries were tested to established the criterion for appraising cryopreservation results. Results The percentage of normal follicles in group B was comparable with group A( P > 0. 05). The value in group C and group D were significant lower than that in group A( P < 0. 05). Quantitative assessment of stromal cell density indicated that the values in group D significantly lower than values in group A while group B and group C had values similar to those of group A. The TUNEL assay showed that the number of positive cells in group A were the lowest of all groups followed by group B,group C and group D( P < 0. 05).The level of BAX transcripts significantly increased in group C and group D( P < 0. 05). The level of CIRP transcripts increased highest in group B followed by group C and group D( P < 0. 05). Conclusion The whole ovary can be appropriately preserved after cryopreservation, and cryoprotectant combination of group B appears to be better for cryopreservation of whole sheep ovary in aspect of histology and anti-apoptosis.
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