Liu Feng-hua, Yang Dong-zi, Wang Yi-feng, Liang Xiao-ping, Peng Wen-ming, Cao Chang-an, Li Qing, Ma Yun, Chen Xi-gu, Guo Zhong-min
Reproductive Medical Center, the Second Peoples Hospital of Guangzhou, Guangzhou, Guangdong 510150, China.
Zhonghua Nan Ke Xue. 2005 Jul;11(7):499-502.
To study transplantation of mouse bone marrow mesenchymal stem cells (MSCs) into the xenogeneic testis.
(1) The tibias and femurs were dissected from 5-6-week-old mice. The marrow in the tibias and femurs was flushed out with medium. MSCs were isolated, cultured and purified in vitro by Percoll density gradient centrifugation combined with adherent method. (2) MSCs of the third generation were adopted and marked with Hoechest33342 for observation, and then made into cell-suspending fluid. (3) The marked MSCs were transplanted into the testis of the xenogeneic mouse by testis net injection. The biopsies of the testis tissues were carried out at different time and made into frost slices at three sites for observation.
(1) A lot of purified MSCs were obtained at the third generation. (2) The nucleoli of the marked MSCs showed light-yellow under the fluoroscope. (3) Xenogeneic transplantation of mouse bone marrow MSCs by testis net injection was successful, without immunoreaction. On the 1 st day after transplantation, MSCs only concentratively distributed in the medial slices, the nucleoli being light-yellow; On the 1 st and 3 rd day, MSCs dispersively distributed in the medial slices; On the 6th, 9th and 12th day, MSCs presented in all the slices of the three sites, some ranging tubally; On the 15th and 18th day, the fluorescence of MSCs weakened; On the 21 st day, the fluorescence of MSCs disappeared.
Transplantation of mouse bone marrow MSCs into the xenogeneic testis by testis net injection is effective and feasible, without immunoreaction. MSCs can survive after transplantation.
研究小鼠骨髓间充质干细胞（MSCs）异体睾丸移植。
（1）取5-6周龄小鼠的胫骨和股骨，用培养基冲洗骨髓，通过Percoll密度梯度离心结合贴壁法体外分离、培养和纯化MSCs。（2）采用第三代MSCs，用Hoechest33342标记后观察，然后制成细胞悬液。（3）将标记的MSCs经睾丸网注射移植到异体小鼠睾丸中，在不同时间取睾丸组织活检，在三个部位制成冰冻切片观察。
（1）第三代获得大量纯化的MSCs。（2）荧光显微镜下标记的MSCs核仁呈浅黄色。（3）经睾丸网注射小鼠骨髓MSCs异体移植成功，无免疫反应。移植后第1天，MSCs仅集中分布在内侧切片，核仁浅黄色；第1天和第3天，MSCs分散分布在内侧切片；第6、9和12天，MSCs出现在三个部位的所有切片中，部分呈管状排列；第15天和第18天，MSCs荧光减弱；第21天，MSCs荧光消失。
经睾丸网注射将小鼠骨髓MSCs移植到异体睾丸中有效可行，无免疫反应，移植后MSCs可存活。
