新生小鼠睾丸组织和作为异种移植物的人类胎儿睾丸组织在免疫缺陷小鼠中的发育情况

摘要

Aim: To investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes. Methods: Immunodeficient nude mice were used as hosts for allografting of neonatal mouse testes and xenografting of human fetal testicular tissues.Stepwise development and stage-specific gene expression of germ cells in allografts were systematically evaluated and parallel compared with those in intact mice by periodically monitoring the graft status with measurement of graft weight, histological analysis and determination of five stage-specific genes. Human testicular tissues from 20 and 26 weeks fetuses were used for the xenografting study. Histological analysis of xenografts was performed 116 and 135 d after the grafting procedure. Results: In the allografting study, progressive increase in tissue volume and weight as well as in tubule diameter in grafts was observed; the appearance time of various germ cells in seminiferous tubules, including spermatogonia, spermatocytes, round and elongate spermatids and sperm, was comparable with that in intact donors; the initiation of gene transcription in grafts showed a similar trend as in normal mice. Graft weight ceased to increase after 7-8 weeks and degradation of grafts was observed after 5 weeks with progressive damage to seminiferous epithelium. In the xenografting study using immature human testicular tissues, graft survival and development was indicated by increasing graft weight, Sertoli cells differentiation into advanced stage, germ cells migration and location to the basal lamina and formation of a niche-like structure. Conclusion: The developmental course and gene expression pattern of germ cells in allografts were similar to those in intact mice. The best time point for retrieval of mouse sperm from grafts was 5-7 weeks after grafting procedure. An accelerated development of immature human testicular cells could be achieved by ectopic xenografting of human testes.%目的:采用同种和异种睾丸组织移植的方法,研究新生小鼠睾丸组织及人类未成熟睾丸组织异种移植物在免疫缺陷小鼠体内发育不同时期生精细胞的组成和基因表达情况中生精细胞的发育情况.方法:以免疫缺陷小鼠为受体,新生小鼠睾丸组织和人类未成熟睾丸组织为供体,分别进行同种和异种移植.通过对移植物的组织形态学观察和分子生物学检测,对各个时期同种移植物中的生精细胞组成及其特异性基因的表达情况进行评估并与未受损小鼠的情况相比较;对人睾丸组织异种移植物的存活及其生精细胞在异体异位的发育情况进行探讨.结果:新生小鼠睾丸组织在成年雄性去势免疫缺陷小鼠体内的发育状况在移植开始的一个阶段与在体睾丸组织的发育情况基本相同,各级生精细胞的出现及其基因表达均与在体睾丸组织中相类似,而移植7-8星期后生精小管发生退化现象.人未成熟睾丸组织在受体中存活并且进一步生长;组织学观察还发现,生精细胞的发育速度与在体相比具有加速的倾向.结论:新生小鼠睾丸组织同种移植物的发育与在体情况基本相同,而人类未成熟睾丸组织异种移植物的发育与正常生理状态相比较呈现出加速的倾向.