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Effects of different freezing methods on the follicular viability of large pieces of human ovarian tissues
ZHAO Shuo ZHAO Shu-qin SHI Qing QIN Xiao-min HAN Yi-long CHAO Lan DENG Xiao-hui*
Acta Anatomica Sinica ›› 2013, Vol. 44 ›› Issue (3) : 427-432.
Effects of different freezing methods on the follicular viability of large pieces of human ovarian tissues
Objective To study effects of cryopreservation of large pieces of human ovarian tissues and assess the follicular
viability after vitrification and two-step freezing. Methods Ovarian tissues from 15 patients were collected. The size of each
tissue block was about 15 mm×15 mm×2mm. The tissue block was cut into three pieces and randomly grouped as fresh(A), vitrification
(B), and two-step freezing (C) groups. All the pieces were cultured in vitro after thawing. The viability and proliferative capacity
of tissues were evaluated by in vitro production of estrogen and progestogen, histological analysis and immunohistochemical
staining of nuclear proliferation-associated antigens, Ki67 and B cell lymphoma/lewkmia-2（Bcl-2）. Results The ratios of normal
follicles were the highest in group A(91.9%), median in group C (82.9%) and the lowest in group B(71.3%) (P<0.05). The ratios of
morphologically normal primordial and primary follicles in group C were 86.8% and 54.4% which were higher than 73.8% and 44.4% in
group B(P<0.05), respectively. Difference of hormonal activity was not found among three groups（P>0.05). Positive expression of
Ki67 of group C（73%）was higher than group B（50%）and group A（55%）(P<0.05), where as that of group A was lower than group B
(P<0.05).There was no difference of positive expression of Bcl-2 staining between group A（57%）and group C（61%）(P>0.05), but
the ratio of positive Bcl-2 expression of group B（42%）was lower than groups A and C (P<0.05). Conclusions The frozen/thawed
large pieces of human ovarian tissues can reserve their vitality and the tissue with two-step freezing is better preserved than
the tissues with vitrification.
Ovarian tissue / Ki67 / Cryopreservation / Vitrification / Two-step freezing / Human
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