人脐血单个核细胞移植治疗放射性裸鼠卵巢早衰

Human Cord Blood Mononuclear Cell Transplantation in the Treatment of Radioactive Experimental Study of Premature Ovarian Failure in Nude Mice

目的:探讨人脐血单个核细胞(HCMNCs)移植治疗卵巢早衰(POF)的可行性及疗效。方法 :雌性BALB/C裸鼠105只,随机分为空白对照组、POF对照组、POF+人脐血单个核细胞移植组。钴60γ射线照射建立POF模型,聚蔗糖(Ficoll)密度梯度离心法制备HCMNCs悬液,并以5-溴-2′-脱氧尿苷(BrdU)标记。POF造模成功后,移植组双侧卵巢内注入标记的HCMNCs悬液,POF对照组双侧卵巢内注入同等量的达尔伯克改良伊格尔氏低培养基(L-DMEM)培养液,空白对照组未行任何处理。移植后4周,酶联免疫吸附试验(ELISA)法测定血清雌二醇(E2)、卵泡刺激素(FSH)、黄体生成激素(LH)水平变化,并取卵巢组织行苏木素-伊红(HE)染色、BrdU组化染色,观察卵巢卵泡数目变化及移植细胞成活情况。结果:在移植组发现移植的脐血单个核细胞可在早衰的卵巢内存活并参与卵巢功能的修复;血清E2、FSH、LH与POF对照组相比,E2明显升高(P<0.05),FSH和LH明显降低(均P<0.05);卵巢切片HE染色示未闭锁卵泡数目有所增加,并可见各个发育阶段卵泡;免疫组化示移植的人脐血单个核细胞可在裸鼠卵巢内定居并存活。结论:HCMNCs可成功植入裸鼠放射性早衰卵巢内,并能改善其功能。

Objective：This study was aimed to explore the feasibility and effects of human cord blood cell transplan- tation for the treatment of premature ovarian failure （POF） in nude mices. Methods：Thirty female nude mouse were ran- domly divided into three groups,Normal control group （n=35） and premature ovarian failure （POF plus vehicle ,n=35） ; POF plus cell transplantation （HCMNCs were implanted into the bilateral ovarian, n=35）. Cobah-60 irradiation was used to establish POF models. HCMNCs were isolated by ficoll density gradient centrifugation ,and labled by BrdU. Four weeks after transplantation, the nude mice were sacrificed for determination of serum E2, FSH, LH,histological and immunochemical ex- amination of the bilateral ovaries. Results ：The transplanted HCMNCs survived in transplantation group and participated the repair process in ovaries. The serums of E2, FSH, LH of the transplantation group were compared with POF control group. E2 significantly increased （ P〈 0.05 ）, FSH decreased significantly （ P〈 0.05 ）, LH decreased significantly （ P〈 0.05 ）. And also found the number of follicles recovered. Conclusions：HCMNCs earl be successfully transplanted into nude mice ovaries, and can improve the recovery of ovarian function.