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用于精原干细胞移植的手术诱导隐睾犬受体模型的建立。

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

作者信息

Lee Won-Young, Lee Ran, Song Hyuk, Hur Tai-Young, Lee Seunghoon, Ahn Jiyun, Jhun Hyunjhung

机构信息

Department of Food Bioscience, RIBHS, College of Biomedical & Health Science, Konkuk University, Chungju, Korea.

Department of Stem Cell and Regenerative Biology, Konkuk University, Seoul, Korea.

出版信息

Lab Anim Res. 2016 Dec;32(4):257-266. doi: 10.5625/lar.2016.32.4.257. Epub 2016 Dec 23.

Abstract

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.

摘要

精原干细胞(SSCs)移植已被用于实验动物模型,以研究生殖系干细胞生物学并培育转基因动物。物种特异性受体模型的制备对于精原干细胞的特性鉴定和后代生产至关重要。在此,我们研究了手术诱导的犬隐睾症作为精原干细胞移植新受体模型的效果。通过手术将睾丸和附睾回纳至腹腔,在十只成年雄性犬中人为诱导单侧或双侧隐睾症。在人为诱导隐睾症(手术)后的一个月内,每周收集睾丸和附睾。为确定手术隐睾症的效果,测量了生精小管直径,并分析了使用PGP9.5和GATA4抗体的免疫组织化学结果。与阴囊内睾丸相比,腹腔内睾丸的生精小管直径显著减小。免疫组织化学结果显示,手术诱导隐睾症后,PGP9.5阳性的未分化精原细胞显著减少,但GATA-4阳性的支持细胞无显著变化。为评估睾丸功能恢复率,则在双侧隐睾症30天后对两只犬进行了睾丸固定术。在睾丸固定术组中,检测到SCP3阳性的精母细胞,并且在睾丸固定术后8周精子发生得以恢复。在本研究中,我们为精原干细胞移植的受体犬模型的手术制备提供了最佳实验条件和时间。此外,我们的数据将有助于通过在非啮齿类动物中使用手术诱导的隐睾症来进行受体准备。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae07/5206233/c224110a0fe1/lar-32-257-g001.jpg

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