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Establishment of a surgically induced cryptorchidism canine recipient model for spermatogonial stem cell transplantation

机译:精原干细胞移植手术诱导隐睾犬受体模型的建立

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摘要

Transplantation of spermatogonial stem cells (SSCs) in experimental animal models has been used to study germ line stem cell biology and to produce transgenic animals. The species-specific recipient model preparation is important for the characterization of SSCs and the production of offspring. Here, we investigated the effects of surgically induced cryptorchidism in dog as a new recipient model for spermatogonial stem cell transplantation. Artificially unilateral or bilateral cryptorchidism was induced in ten mature male dogs by surgically returning the testis and epididymis to the abdominal cavity. The testes and epididymides were collected every week after the induction of artificial cryptorchidism (surgery) for one month. To determine the effect of surgical cryptorchidism, the seminiferous tubule diameter was measured and immunohistochemistry using PGP9.5 and GATA4 antibodies was analyzed. The diameters of the seminiferous tubules of abdominal testes were significantly reduced compared to those of the scrotal testes. Immunohistochemistry results showed that PGP9.5 positive undifferentiated spermatogonia were significantly reduced after surgical cryptorchidism induction, but there were no significant changes in GATA-4 positive sertoli cells. To evaluate the testis function recovery rate, orchiopexy was performed on two dogs after 30 days of bilateral cryptorchidism. In the orchiopexy group, SCP3 positive spermatocytes were detected, and spermatogenesis was recovered 8 weeks after orchiopexy. In this study, we provided optimum experimental conditions and time for surgical preparation of a recipient canine model for SSC transplantation. Additionally, our data will contribute to recipient preparation by using surgically induced cryptorchidism in non-rodent species.
机译:实验动物模型中精原干细胞(SSC)的移植已用于研究生殖系干细胞生物学并生产转基因动物。特定物种的受体模型准备对于SSC的表征和后代的生产很重要。在这里,我们调查了狗手术诱导的隐睾症作为精原干细胞移植的新受体模型的影响。通过手术将睾丸和附睾返回腹腔,在十只成熟的雄性狗中诱发了人为的单侧或双侧隐睾症。人工隐睾症(手术)诱导一个月后,每周收集一次睾丸和附睾。为了确定手术隐睾症的效果,测量了生精小管的直径,并分析了使用PGP9.5和GATA4抗体的免疫组织化学。与阴囊睾丸相比,腹部睾丸生精管的直径明显减小。免疫组织化学结果显示,经手术隐睾诱导后,PGP9.5阳性未分化精原细胞明显减少,但GATA-4阳性睾丸支持细胞无明显变化。为了评估睾丸功能恢复率,在双侧隐睾症治疗30天后对两只狗进行了睾丸检查。睾丸切除术组检测到SCP3阳性精细胞,并在睾丸切除术后8周恢复精子生成。在这项研究中,我们为SSC移植的犬科动物模型的外科手术准备提供了最佳的实验条件和时间。此外,我们的数据将通过在非啮齿类动物中使用手术诱发的隐睾症而有助于接受者的准备。

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