The feasibility study on human ovarian tissue vitrification using self-made cryoloop.

LI Yu-feng; L(U) Yan; LIU Qun; REN Xin-ling; HU Juan; LIU Xing; ZHANG Han-wang; LIU Cong

Abstract：Objective To investigate cryopreservation effects of human ovarian tissue using self-made cryoloop. Methods Ovarian biopsies collected from 5 women of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from May to September in 2008 were vitrificated by cryoprotectants, composed of ethylene gly-col, dimethylsuphoxide and sucrose, using self-made cryoloop as a cooling device. The morphology analysis of primordial and primary follicles,apoptosis signals, ultastructure,endocriological function cultured in vitro of the fresh and vitrificat-ed/thawed ovarian tissues were accessed. Results The ratio of morphologically normal primordial and primary follicle(84.47±4.66)was not different compared with the fresh one(89.46±4.94), P>0.05. The mean TUNEL-pssitive sur-face area values of the fresh tissue and the vitrifieated one were (0.07±0.02) %, (0.10±0.05)%, and no statistically significant difference was found(P>0.05).The uhrastructure was similarly preserved after vitrification. After culture of fresh tissue pieces and vitrifieated/thawed pieces, the supernatants showed average estrodiol concentration of (2549. 73 ±711. 87 ) pmoL/L, ( 2514. 87±714. 66)pmol/L,respectively, and no statistically significant difference was found (P>0.05). Conclusion Cryolcop vitrification is an effective, simple and inexpensive alternative for human ovarian tissue cryopreservation.