Tian Ru-hui, Hu Hong-liang, Liu Ping, Li Peng, Yang Shi, Zhu Yong, Ma Meng, Sun Can, Zou Sha-sha, Guo Xi-zhi, Huang Yi-ran, Li Zheng
Department of Urology/Laboratory of Sperm Development and Genetics, Shanghai Human Sperm Bank, Shanghai Institute of Andrology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200001, China.
Zhonghua Nan Ke Xue. 2011 Oct;17(10):867-72.
To investigate the ectopic grafts of mouse testicular cells by observing the reconstruction of seminiferous tubules, colonization of spermatogenic cells and spermatogenesis using immunodeficient mice as recipients.
The testes of newborn male ICR mice were digested to obtain single cell suspension. The cells were then mixed with matrigel and subcutaneously grafted into the dorsal region of the male nude mice. The mice were castrated after the operation and the grafts were dissected from 5 of the nude mice at 4, 6, 8 and 10 weeks, respectively. The success rates of transplantation and the graft diameters were calculated, and the structure of the reconstituted seminiferous tubules, colonization of the germ cells and spermatogenesis were observed by HE staining and immunohistochemistry.
All the mice recipients survived after the testicular cell transplantation. Within 10 weeks after the operation, tissue masses could be observed, with the diameter increased from (3.91 +/- 0.71) mm at 4 weeks to (6.69 +/- 0.50) mm. Neovascularization was detected at the surface of the masses and seminiferous tubule structures found in the grafts. The germ cells that developed from spermatogonia to round spermatids were observed, but with no sperm in the tubules. Germ cells, Sertoli cells and Leydig cells were identified by immunochemical detection of Mvh, Gata4 and P450Scc in the grafts at 8 weeks.
Seminiferous tubules could be ectopically reconstructed from suspension of neonatal mouse testicular cells. Ectopic grafting provided a preferable model for the studies on testis tissue engineering and interactions between testicular cells during testicular development and spermatogenesis.
以免疫缺陷小鼠为受体,通过观察生精小管的重建、生精细胞的定植和精子发生情况,研究小鼠睾丸细胞的异位移植。
消化新生雄性ICR小鼠的睾丸以获得单细胞悬液。然后将细胞与基质胶混合,皮下移植到雄性裸鼠的背部区域。术后对小鼠进行去势,并分别在4、6、8和10周时从5只裸鼠中取出移植体。计算移植成功率和移植体直径,通过苏木精-伊红染色和免疫组织化学观察重建生精小管的结构、生殖细胞的定植和精子发生情况。
睾丸细胞移植后所有受体小鼠均存活。术后10周内可观察到组织块,直径从4周时的(3.91±0.71)mm增加到(6.69±0.50)mm。在组织块表面检测到新生血管,移植体中发现生精小管结构。观察到从精原细胞发育为圆形精子细胞的生殖细胞,但小管内无精子。在8周时通过对移植体中Mvh、Gata4和P450Scc进行免疫化学检测鉴定出生殖细胞、支持细胞和间质细胞。
新生小鼠睾丸细胞悬液可异位重建生精小管。异位移植为睾丸组织工程以及睾丸发育和精子发生过程中睾丸细胞间相互作用的研究提供了一个较好的模型。