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Background: The positive effect of cooling on tissue cells is known. The aim of this research was to study the intensiveness of neo-vascularisation and follicular development in ovarian tissue after 24 hours cooling to 5°C before cryopreservation.
Methods: Fifty six pieces from 7 patients were divided into the following four groups: Group 1: pieces cultured just after operation, Group 2: pieces cooled after operation to 5°C for 24 hours and then cultured, Group 3: pieces frozen-thawed just after operation and then cultured, Group 4: pieces cooled after operation to 5°C for 24 hours, frozen, thawed, and then cultured. Culture of ovarian pieces was performed in a chorioallantoic membrane (CAM)-system for 5 days. The efficacy of the tissue cooling was evaluated by the development of follicles and intensiveness of neo-vascularisation (by Desmin).
Results: For Group 1, 2, 3, and 4, mean density of follicles per 1 mm3 was 10.1, 11.1, 9.8, and 12.0, respectively (p1-2, 3-4 < 0.05). For these groups 91%, 92%, 90%, and 90% preantral follicles were morphologically normal (p1-2, 3-4 > 0.1). The immunohistochemical analysis showed that the intensiveness of neo-vascularisation observed in ovarian tissue of Group 2 (pre-cooled before culture) and Group 4 (pre-cooled before cryopreservation) was drastically increased.
Conclusions: The 24 hour cooling to 5°C before cryopreservation is beneficial for cryopreservation of human ovarian tissue.
DOI: 10.7754/Clin.Lab.2012.120420
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