] 目的 探讨莱迪希细胞（Leydig cell, LC）和谢尔托利细胞（Sertoli cell, SC）共移植构建的组织工程化雄激素分泌组织同种异基因移植的可行性。并在此基础 上，进一步建立分离、培养人睾丸组织 LC 及 SC 的技术体系。方法 对应用胶原酶消 化及差速贴壁法从同一鼠、人睾丸组织中分离、培养的 LC 和 SC 进行鉴定及纯度分析。 LC 及 SC 与聚乳酸（Polylactic acid, PLA）/聚乙醇酸（Polyglycolic acid, PGA）生物材 料复合后，移植于去势的同基因和同种异基因大鼠体内，定期检测两组动物体内的血 睾酮水平。结果 应用酶消化和差速贴壁法可以从同一大鼠、人睾丸组织中，获得数 量充足、纯度和功能良好的 LC 及 SC。LC 与 SC 共移植于去势的同基因和同种异基因 大鼠体内可以形成具有分泌雄激素功能的均质实体组织，激素分泌可持续三个月以 上。结论 应用酶消化和差速贴壁法可以从大鼠和人的睾丸组织内获得数量充足、功 能良好的 LC 和 SC；LC 与 SC 共移植可以作为组织工程化雄激素分泌组织构建的良好 种子细胞；构建的组织工程化雄激素分泌组织具有同种异基因移植的可行性（≥3 个 月）。 [关键词] 组织工程；睾丸；间质细胞；支持细胞；雄激素；免疫豁免；同种异基因 移植 Feasibility Study on Allogenic Transplantation of Tissue Engineered Testeosterone-Secreting Tissue [abstract] Objective To investigate the feasibility on allogenic transplantation of tissue engineered testeosterone-secreting tissue with co-transplanted LC and SC. Furthermore, establish an isolation and culture system of human testicular LC and SC on the foundation of this technique. Methods LC and SC were got by collagenase digestion and different-speed adherence methods from the same rat or human testis, and the characteristic and purity of the isolated LC and SC were identified and analyzed. The LC and SC were seeded onto the PGA/PLA material to form cell-scaffold constructs. The cell-scaffoldconstructs were transplanted into the gastrocolic omentum of the castrated homogeneic or allogeneic rats. The serum testosterone was assayed at regular intervals. Results Both LC and SC can be got from the same rat or human testis by collagenase digestion and different-speed adherence methods with high purity and good function. And after isotransplantated or allotransplantated into the castrated rats, the secreting function of the constructs could last at least 3 month. Conclusion Abundant LC and SC with high pure and good function can be got by collagenase digestion and different-speed adherence methods from the same rat or human testis. It is feasible that to construst the tissue engineered testeosterone-secreting tissue with co-transplantation LC and SC in allogeneic individuals. [key words] tissue enginnering; testis; Leydig cells; Sertoli cells; androgen; immune privilege；allogenic transplantation第二军医大学博士学位报告 目录 整形外科 目 录 缩略词表 ….1 中文摘要.3 英文摘要 ….6 前 言.9 第一部分 种子细胞分离培养体系及雄激素分泌组织构建技术优化…..….14 实验一 大鼠睾丸 LC、SC 分离培养及其基本生物学特性研究.…..14 材料和方法...14 结果…...28 讨论…...34 实验二 “细胞-生物材料”复合物的形成及同基因移植后的初步鉴定.….38 材料和方法...38 结果…...44 讨论…...49 第二部分 组织工程化雄激素分泌组织同种异基因移植可行性初步研究...51 材料和方法...51 结果…...55 讨论…...67 第三部分 人睾丸LC、SC分离培养及其基本生物学特性研究…..72 材料和方法...72 结果.…..77 讨论…...83 全文总结.…..86