Wang Xiang, Ding Qiang, Zhang Yuanfang, Wang Huilin, Ma Lianghong, Xie Xiayang
Department of Urology, Huashan Hospital, Fudan University, Shanghai, P. R. China.
Fertil Steril. 2008 Oct;90(4 Suppl):1538-49. doi: 10.1016/j.fertnstert.2007.08.049. Epub 2008 Jan 3.
To evaluate restoration of spermatogenesis and fertility of the recipients who underwent high-dose ablative treatment before freeze-thawed spermatogonial stem cell (SSC) allotransplantation.
Prospective experimental study.
University-based teaching hospital.
ANIMAL(S): Adult busulfan-treated BALB/c nude mice as recipients of C(57)BL/6 pup SSCs, oocytes of 4-to 6-week-old F(1) hybrid mice for IVF, adult closed group ICR mouse as pseudopregnant female.
INTERVENTION(S): Isolation, purification, and fresh and freeze-thawed transplantation, IVF, embryo transplantation.
MAIN OUTCOME MEASURE(S): Cell viability, Western blot, real-time fluorescence quantitative-polymerase chain reaction (PCR), scanning electron microscope (SEM), outcome of IVF, and embryo transplantation.
RESULT(S): Western blot, real-time fluorescence quantitative-PCR, and SEM revealed different spermatogenesis where the fresh groups were uniquely higher than the freeze-thawed groups. A total of six developed two-cell embryos obtained by IVF were transplanted into the oviduct of a pseudopregnant ICR mouse, which resulted in a birth of two paternally different pups. Two female offspring grew into healthy adults with no apparent abnormalities.
CONCLUSION(S): The results provide the first solid evidence that spermatozoa generated from the busulfan-treated recipient and donor cryopreserved SSCs were both fertile. Ultimate evaluation of the SSC cryopreservation and transplantation technology rests on their functional capacity to generate desirable donor spermatogenesis and fertility in the recipient.
评估在冻融精原干细胞（SSC）同种异体移植前接受高剂量消融治疗的受体的精子发生恢复情况和生育能力。
前瞻性实验研究。
大学附属医院。
经白消安处理的成年BALB/c裸鼠作为C57BL/6幼鼠SSC的受体，4至6周龄F1杂交小鼠的卵母细胞用于体外受精，成年封闭群ICR小鼠作为假孕雌性。
分离、纯化、新鲜和冻融移植、体外受精、胚胎移植。
细胞活力、蛋白质免疫印迹法、实时荧光定量聚合酶链反应（PCR）、扫描电子显微镜（SEM）、体外受精结果和胚胎移植。
蛋白质免疫印迹法、实时荧光定量PCR和SEM显示不同的精子发生情况，其中新鲜组明显高于冻融组。通过体外受精获得的总共6个发育到二细胞期的胚胎被移植到一只假孕ICR小鼠的输卵管中，结果产下了两只父系不同的幼崽。两只雌性后代成长为健康的成年个体，无明显异常。
结果首次提供了确凿证据，表明经白消安处理的受体和供体冷冻保存的SSC产生的精子均具有生育能力。对SSC冷冻保存和移植技术的最终评估取决于它们在受体中产生理想的供体精子发生和生育能力的功能。
