Ren Shang-Qing, Deng Shi, Wu Yun-Jian, Hou Lin, Dong Tian, Dong Qiang
Sichuan Da Xue Xue Bao Yi Xue Ban. 2014 Mar;45(2):192-5, 201.
To study the differentiation of human bone marrow mesenchymal stem cells (BMSCs) into Leydig or steroidogenic cells in vivo and the immunoreaction related to transplantation into mouse testis.
After differentiation and cultivation, the 3rd-passage BMSCs were collected and labeled with Hoechest 33342, and joined the saline fluorouracil to form cell-suspending fluid. After injection of the etgane dimethane sulphonate (EDS), the mice received the transplanted cell-suspending fluid by testis net injection with a dose of each side testicular 0.05 mL. Since the first day prior to transplantation, mice were executed every 2 d (one mouse each time) and the testosterone concentrations were analyzed. The control group included 20 BALB/c mice without any treatment during the same period. The results were analyzed by microscopic observation, using 3beta-hydroxysteroid dehydrogenase (3beta-HSD) monoclonal antibody and mouse anti-human cell nucleus monoclonal antibody for immunofluorescence assay on the testis. For tracking the BMSCs, the cells which were positively stained with both 3beta-HSD and mouse anti-human cell nucleus monoclonal antibodies were retrieved.
Certain killing effect of EDS to the mouse Leydig cells was observed. Transplantation of human BMSCs into the mouse testis by testis net injection was effective and feasible, no immunoreactions were detected. After transplantation, no positive cells of 3beta-HSD) and mouse anti-human cell nucleus monoclonal antibody were found.
Transplantation of human BMSCs into the mouse testis by testis net injection was effective and feasible, no immunoreactions were detected. After transplantation, the human BMSCs failed to differentiate into Leydig cells or steroidogenic cells.
研究人骨髓间充质干细胞（BMSCs）在体内向睾丸间质细胞或类固醇生成细胞的分化情况以及与移植入小鼠睾丸相关的免疫反应。
分化培养后，收集第3代BMSCs，用Hoechest 33342标记，加入生理盐水氟尿嘧啶制成细胞悬液。注射乙烯雌酚二甲磺酸盐（EDS）后，通过睾丸网注射将细胞悬液移植到小鼠体内，每侧睾丸注射剂量为0.05 mL。自移植前一天起，每2天处死1只小鼠（每次1只），分析睾酮浓度。对照组为20只同期未作任何处理的BALB/c小鼠。通过显微镜观察、用3β-羟基类固醇脱氢酶（3β-HSD）单克隆抗体和小鼠抗人细胞核单克隆抗体对睾丸进行免疫荧光分析来分析结果。为追踪BMSCs，检索同时被3β-HSD和小鼠抗人细胞核单克隆抗体阳性染色的细胞。
观察到EDS对小鼠睾丸间质细胞有一定杀伤作用。通过睾丸网注射将人BMSCs移植到小鼠睾丸有效可行，未检测到免疫反应。移植后，未发现3β-HSD和小鼠抗人细胞核单克隆抗体的阳性细胞。
通过睾丸网注射将人BMSCs移植到小鼠睾丸有效可行，未检测到免疫反应。移植后，人BMSCs未能分化为睾丸间质细胞或类固醇生成细胞。
