Maltaris Theodoros, Dimmler Arno, Müller Andreas, Hoffmann Inge, Beckmann Matthias W, Dittrich Ralf
Department of Obstetrics and Gynecology, Erlangen University Hospital, Erlangen, Germany.
J Obstet Gynaecol Res. 2006 Jun;32(3):273-9. doi: 10.1111/j.1447-0756.2006.00398.x.
To compare two freezing protocols in an automatic open-vessel freezing system for cryopreservation of rat ovarian tissue.
Ovarian tissue was transplanted heterotopically into the neck muscle, either without cryopreservation (group 1, n = 6) or with cryopreservation after equilibration with 1.5 mol/L dimethyl sulfoxide and propanediol (protocol A, group 2, n = 6) or 1.5 mol/L ethyl glycol (protocol B, group 3, n = 6). The ovarian tissue was examined with LIVE/DEAD fluorescent viability staining and histologically after isotransplantation.
The healthy follicular loss (intact oocyte and >50% granulosa cells alive) due to cryopreservation was 15.5% with protocol A and 12.2% with protocol B. Histological examination showed follicles in all developmental phases in all groups: group 1, 35.5 +/- 5.7/mm(2) (mean +/- SD); group 2, 16.0 +/- 5.0/mm(2); group 3, 17.3 +/- 5.7/mm(2). The differences between groups 1 and 2 and between groups 1 and 3 were significant (P < 0.001). The difference between groups 2 and 3 was not significant (P = 0.33).
These results demonstrate that the use of an open freezing system with both freezing protocols allows cryopreservation of rat ovarian tissue with equally good survival rates.
比较自动开放式容器冷冻系统中两种冷冻方案用于大鼠卵巢组织冷冻保存的效果。
将卵巢组织异位移植到颈部肌肉，一组不进行冷冻保存（第1组，n = 6），另外两组在与1.5摩尔/升二甲基亚砜和丙二醇平衡后进行冷冻保存（方案A，第2组，n = 6）或与1.5摩尔/升乙二醇平衡后进行冷冻保存（方案B，第3组，n = 6）。同种移植后，用LIVE/DEAD荧光活力染色和组织学方法检查卵巢组织。
方案A冷冻保存导致的健康卵泡损失（完整卵母细胞和>50%颗粒细胞存活）为15.5%，方案B为12.2%。组织学检查显示所有组均有各发育阶段的卵泡：第1组，35.5±5.7个/mm²（平均值±标准差）；第2组，16.0±5.0个/mm²；第3组，17.3±5.7个/mm²。第1组与第2组之间以及第1组与第3组之间的差异有统计学意义（P < 0.001）。第2组与第3组之间的差异无统计学意义（P = 0.33）。
这些结果表明，使用两种冷冻方案的开放式冷冻系统均可实现大鼠卵巢组织的冷冻保存，且存活率相当。
