小鼠心脏移植保存期间经体外灌注行重组腺病毒基因转染的研究

目的:探讨小鼠心脏移植中,通过在体外保存期间对供心进行灌注,进行重组腺病毒基因转染的方法及效率。方法: 利用小鼠异位心脏移植,在供心保存期间行重组腺病毒灌注,在移植后7 d检测移植物内标记基因的表达情况,并通过免疫组织化学染色检测移植物内免疫细胞的浸润,通过流式细胞仪检测受体淋巴细胞的激活状态。结果: 小鼠供心通过体外灌注进行重组腺病毒基因转导,并在4 ℃保存2 h,复跳率达100%,移植后1周,标记基因表达强度到达峰值。移植物组织结构完整,无明显细胞浸润。受体淋巴细胞的激活状态与空白对照组相比,无明显差异。结论: 小鼠心脏移植保存期间体外灌注可应用于腺病毒基因转染等移植物处理过程。

In vitro perfusion method during cardiac graft preservation and its application in adenovirus mediated gene transfection

AIM: To establish a method of in vitro donor heart perfusion in murine cardiac transplantation during preservation and apply it in adenovirus mediated gene transfection for donor heart. METHODS: Donor heart was transfected with recombinant adenovirus and stored for 2 hours after harvest, then it was transplanted heterotopically in abdomen. The grafts were appraisal by palpitation. Marked gene products were determined by X-Gal staining, aod T cell infiltration was determined by immunohistochemistry. The activation markers of recipients' lymphocytes were examed by cytometry. RESULTS: The grafts survival rate is 100% after perfusion and cold storage. The LacZ staining became strong 1 week after transplantation. The grafts remained an intact structure and no apparent T cell infiltration. The activation status of recipients' lymphocytes were not enhanced by transfected cardiac graft. CONCLUSION: In vitro perfusion during graft cold preservation is feasible for adenovirus mediated gene transfection. [