小鼠睾丸消化细胞异体移植重构生精小管的研究  

Reconstructed seminiferous tubules from ectopic grafting of testicular cells

作　　者：田汝辉[1,2,3] 胡洪亮[1,2,3] 刘平[1,2,3] 李朋[1,2,3] 杨施[1,2,3] 朱勇[1,2,3] 马猛[1,2,3] 孙璨[1,2,3] 邹沙沙[1,2,3] 郭熙志[4] 黄翼然[1,2,3] 李铮[1,2,3] 

机构地区：[1]上海交通大学医学院附属仁济医院泌尿外科 [2]上海市男科学研究所精子发育与遗传学实验室 [3]上海市人类精子库,上海200001 [4]上海交通大学Bio-X中心,上海200240

出　　处：《中华男科学杂志》2011年第10期867-872,共6页National Journal of Andrology

基　　金：国家重点基础研究发展计划(2010CB945200;2011CB944504);上海市教委重点课题(10ZZ70);上海交通大学自然基金课题(2008XJ017)~~

摘　　要：目的:采用免疫缺陷小鼠作为受体,通过对小鼠睾丸消化细胞异位移植后不同时期移植物的研究,观察生精小管重构、生精细胞归巢及精子发生情况。方法:取新生ICR小鼠的睾丸消化成单细胞悬液,将其与Matrigel基质胶混匀后移植于雄性裸鼠背部皮下,术后裸鼠行去势。移植后分别于4、6、8、10周处死5只裸鼠,计算移植成功率,取移植物测量直径,并进行HE染色和免疫组化检测,观察生精小管的重构、生精细胞归巢及精子发生情况。结果:20只受体鼠接受睾丸消化细胞移植后全部存活。睾丸消化细胞移植后10周内可见明显隆起的包块,包块直径由第4周的(3.91±0.71)mm增加到(6.69±0.50)mm,移植物表面有血管生成。对移植物石蜡切片进行HE染色可见生精小管样结构,部分生精小管管腔内可见由精原细胞发育至精子细胞的各级生殖细胞,未见明显精子产生。对8周移植物进行免疫组化观察,可见生殖细胞标志物Mvh、支持细胞标志物Gata4和间质细胞标志物P450Scc表达。结论:新生小鼠睾丸消化细胞移植于裸鼠背部皮下后可重构生精小管,为研究睾丸组织工程及睾丸发育和精子发生过程中睾丸各组成细胞之间的相互作用提供了理想的研究模型。Objective： 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.Methods： 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.Results： 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.Conclusion： 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.

关 键 词：睾丸消化细胞 异位重构 生精小管 小鼠 

分 类 号：R114[医药卫生—卫生毒理学]