摘要
背景:精原干细胞作为精子发生过程的基础和前提,其自我更新和分化途径目前仍不完全清楚。目的:观察非免疫缺陷动物新生Wistar大鼠的精原干细胞和睾丸组织移植于去势成年Wistar大鼠后的成活及生长发育情况。设计、时间及地点:组织细胞形态学水平的随机动物对照实验,于2007-04/08在广西医科大学实验动物中心外科实验室完成。材料:选用健康新生7～9d雄性Wistar大鼠为供体,受体为经过严格检疫合格的8～12周的成年雄性Wistar大鼠,体质量180～220g。方法:取新生雄性大鼠睾丸,采用两步法组合酶顺序消化制备大鼠精原干细胞悬液,以Percoll不连续密度梯度离心法初步纯化精原干细胞。取10只成年雄性大鼠,切除双侧睾丸形成去势大鼠,按随机数字表法分为2组,每组5只。精原干细胞悬液移植组将制取好的1mL精原干细胞悬液在5min内注射至受体背部皮下。睾丸组织块移植组将制备好的已剖开的睾丸组织植入受体背部皮下,每个受体移植2个睾丸4块睾丸组织。主要观察指标:移植物的生长发育情况,移植8周末移植物的组织学特点及C-kit免疫组化定性分析结果。结果:精原干细胞悬液移植后,移植物未见成活生长。睾丸组织块移植后移植物部分成活,移植8周末组织学检查可见特征性的精曲小管和细胞结构,可见精子细胞,部分生精小管退化;睾丸间质中可见淋巴细胞浸润;免疫组化鉴定可见睾丸组织内C-kit阳性细胞表达。结论:同种异体异位移植于非免疫缺陷鼠中,新生睾丸组织块可以成活并能形成精子细胞,而精原干细胞悬液移植后未见移植物生长。
BACKGROUND： The spermatogonial stem cell （SSc） is the premise of spermatogenesis process, of which the self-renewal and differentiation pathway remains unclear. OBJECTIVE： To explore the growth and development of allograft of SSc and testis tissue grafted to the non-immunodeficiency Wistar rats and ovariectomized Wistar rats. DESIGN, TIME AND SETTING： The randomized control experiment on cellular morphology was conducted in the Surgical Laboratory of Animal Experiment Center in Guangxi Medical University from April to August 2007. MATERIALS： Neonatal Wister rats with 7 9 days old were served as donors and adult male Wister rats （8-12 weeks）, with clear grade, weighting 180 220g, were selected as recipients. METHODS： The testis were aseptically isolated from neonatal Wister rats, and prepared for SSc suspension by two-step enzymatic digestion. The Percoll discontinuous density gradient method was used to purify SSc. 10 adult male rats, after performed gonadectomy, were divided into 2 groups by random number table. Animal in the SSc group were injected 1 mL SSc suspension into skin of the back, while testis and testis tissues were grafted under skin in the other group. MAIN OUTCOME MEASURES： The growth and development of grafts, histological characteristics, as well as qualitative analysis results of C-kit immunohistochemistry were observed. RESULTS： The allograft of SSc as recepients could not survive and grow. The grafted testis tissues could partly survive in the testis group. Histology observation showed that the seminiferous tubules, spermatogonial cell could be found, partly of seminiferous tubule had degenerated, and a lot of lymphocyte infilitration in testis interstitial tissue. Positive expression of C-kit ceils could be seen in testis tissues. CONCLUSION： After allotransplantation on non-immunodeficiency rats, the grafted testis tissue can survival and develop to spermatid. However, this can not been seen in the SSc group.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第B12期10411-10414,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广西卫生厅基金资助课题(桂卫E2002029)"同种异体原始生殖细胞移植的研究"~~
