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Hum. Reprod. Advance Access published online on March 10, 2005
Human Reproduction, doi:10.1093/humrep/deh787
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Human Reproduction © European Society of Human Reproduction and Embryology 2005; all rights reserved
Received January 5, 2004
Revised November 17, 2004
Accepted January 13, 2005

Article

Effects of mouse ovarian tissue cryopreservation on granulosa cell-oocyte interaction

P. Navarro-Costa 1, S.C. Correia 2, A. Gouveia-Oliveira 3, F. Negreiro 3, S. Jorge 4, A.J. Cidadão 1, M.J. Carvalho 2, and C.E. Plancha 1*

1 Biology of Reproduction Unit, Institute of Molecular Medicine, Lisbon Medical School, 1649-028 Lisboa, Portugal
2 Medicine of Reproduction Unit, Gynaecology Service, Maternidade Dr. Alfredo da Costa, 1069-089 Lisboa, Portugal
3 Datamedica Lda., Rua Rosa Araújo, 34, 5°, 1250-195 Lisboa, Portugal
4 Medicine of Reproduction Unit, Gynaecology Service, Maternidade Dr. Alfredo da Costa, 1069-089 Lisboa, Portugal

* To whom correspondence should be addressed.
C.E. Plancha, E-mail: carlos.plancha{at}mail.telepac.pt


  Abstract

BACKGROUND: Current ovarian tissue cryopreservation protocols have yet to be assessed in terms of somatic-germ cell interaction. Accordingly, post-thaw analysis of antral follicles can yield relevant data on the disruption of the granulosa-oocyte interface. METHODS: We compared fresh mouse ovarian tissue with tissues that had been either cryopreserved using dimethylsulphoxide (DMSO) or glycerol as cryoprotectants, or exposed to such cryoprotectants without freezing. The assessed parameters were: number of immature oocytes retrieved per ovary, allocation of the oocytes to different classes regarding antral follicle size and oocyte-granulosa cell adhesion, and the relative density of transzonal processes containing filamentous actin (TZPs-Act). RESULTS: Although cryopreservation reduces the average number of oocytes retrieved per ovary, it increases the relative distribution of granulosa-free oocytes while decreasing that of granulosa-enclosed ones. Additionally, a post-thaw decrease in TZPs-Act density was recorded. This decrease was also observed after cryoprotectant exposure without freezing, although at a lower level. For the assessed parameters, DMSO was more effective than glycerol as a cryoprotectant. CONCLUSIONS: In situ cryopreservation of granulosa-oocyte complexes with current protocols disrupts the granulosa-oocyte interface. The different patterns of granulosa cell adhesion and interaction in oocytes derived from different-sized antral follicles further suggests that the granulosa-oocyte interface may be developmentally regulated.

Keywords: actin; granulosa-oocyte communication; ovarian tissue cryopreservation; transzonal processes.
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[Abstract] [Full Text] [PDF]


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