【摘要】 目的:研究小鼠骨髓间充质干细胞(MSCs)移植异体睾丸。方法:①分离5~6周龄的BALB/c小鼠胫骨、股骨,冲出骨髓,用Percoll密度梯度离心法和贴壁法分离、培养、纯化MSCs。②取第3代MSCs,用Hoe-chest33342标记并观察,制备细胞悬液。③用睾丸网注射法将标记的MSCs移植入异体小鼠睾丸内,于各时相点取睾丸组织,在3个位点连续制作快速冰冻切片,镜下观察。结果:①传至3代即可得到数量较多的纯化的MSCs。②在荧光显微镜下,标记的MSCs细胞核呈亮黄色。③用睾丸网注射法行小鼠MSCs移植异体睾丸成功,未见免疫排斥反应发生。移植后1、3d,只在中间的切片上见到MSCs,细胞核呈亮黄色,两侧的切片未查到MSCs,移植后1d细胞较集中,移植后3d细胞散在分布;移植后6、9、12d,3个位点的切片均见MSCs,部分MSCs排列呈管状;移植后15、18d,MSCs的核荧光有所减弱;移植后21d,荧光消失。结论:用睾丸网注射法将小鼠MSCs移植异体睾丸获得成功,未见免疫排斥反应发生,移植后的MSCs存活。更多还原

【Abstract】 Objective: To study transplantation of mouse bone marrow mesenchymal stem cells(MSCs) into the xenogeneic testis.Methods: 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. MSCs of the third generation were adopted and marked with Hoechest33342 for observation, and then made into cell-suspending fluid. 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.Results: A lot of purified MSCs were obtained at the third generation. The nucleoli of the marked MSCs showed light-yellow under the fluoroscope. 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 6(th), 9(th)and 12(th) day, MSCs presented in all the slices of the three sites, some ranging tubally; On the 15(th) and 18(th) day, the fluorescence of MSCs weakened; On the 21(st) day, the fluorescence of MSCs disappeared.Conclusion: 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.更多还原