【摘要】：目的:探討適合人卵巢組織玻璃化冷凍的冷凍保護劑及合適載體。方法:收集40例因卵巢良性包塊行手術者的卵巢皮質40片,將每片卵巢皮質切為20塊卵巢組織塊。將其隨機分為10組,玻璃化冷凍組9組,另一組為對照組,9組玻璃化冷凍組再根據使用不同配伍濃度的冷凍保護劑(CPA1、CPA2、CPA3)及3種不同載體方法(微滴法、凍存管法、篩網法)進行細分。分析比較玻璃化冷凍組9個亞組與對照組凍融前后卵泡形態(tài)學變化、細胞增殖及凋亡程度。結果:1形態(tài)學HE染色示:僅CPA1組微滴法和篩網法、CPA2組微滴法與對照組比較,差異無統(tǒng)計學意義(P0.05)。2免疫組化示:僅CPA1組微滴法和篩網法卵巢組織卵泡的Ki-67表達與對照組比較,差異無統(tǒng)計學意義(P0.05)。3Tunel細胞凋亡檢測示:各級卵泡中卵母細胞及顆粒細胞中均未見凋亡細胞。結論:低濃度冷凍保護劑DMSO聯合EG(即CPA1液)使用微滴法或篩網為冷凍載體凍存卵巢組織,對卵巢組織的損傷最小。
[Abstract]:Objective: to study the suitable cryopreservation agent and carrier for vitrification of human ovarian tissue. Methods: 40 pieces of ovarian cortex were collected from 40 patients with benign ovarian mass. Each ovarian cortex was cut into 20 ovarian tissue blocks. They were randomly divided into 10 groups: vitrification group (n = 9) and control group (n = 9). According to the use of CPA1,CPA2,CPA3 and three different carrier methods (microdrop method, cryopreservation method), the vitrification group was treated with different concentrations of cryopreservation agent (CPA1,CPA2,CPA3). Screen method) for subdivision. The morphological changes, proliferation and apoptosis of follicles in 9 subgroups of vitrification group and control group were analyzed and compared before and after freezing and thawing. Results: 1Morphologic HE staining showed that only CPA1 group and sieve method were used, and CPA2 group was compared with control group. There was no significant difference (P0.05). 2 Immunohistochemistry showed that the expression of Ki-67 in ovarian follicles of CPA1 group was higher than that of control group. There was no significant difference (P0.05). Apoptosis of 3Tunel cells was not found in oocytes and granulosa cells in all follicles. Conclusion: cryopreservation of ovarian tissue with low concentration DMSO combined with EG (CPA1 solution) using microdrop method or sieve as cryopreservation carrier can cause the least damage to ovarian tissue.
【作者單位】： 天津醫(yī)科大學研究生院;南開大學;天津市中心婦產科醫(yī)院;
【基金】：天津市衛(wèi)生局資助項目(編號:12ZK101)
【分類號】：R714.8

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