Cardoso M T, Pinheiro A O, Vidane A S, Casals J B, de Oliveira V C, Gonçalves Njn, Martins D S, Ambrósio C E
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of Sao Paulo, Pirassununga, SP, Brazil.
Department of Surgery, Faculty of Veterinary Medicine and Animal Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.
Reprod Domest Anim. 2017 Apr;52 Suppl 2:58-64. doi: 10.1111/rda.12832. Epub 2016 Oct 23.
The biosafety of innovative procedures that utilize stem cells in regenerative medicine has been addressed in several studies. Previous work has showed no tumour formation following the use of feline and human amniotic membrane-derived stem cells (AMSCs). In contrast, tumour formation was observed when canine AMSCs were utilized. These findings suggested that feline and human, but not canine, AMSCs are suitable for cell transplantation trials. This study aimed to further evaluate the feasibility of utilizing canine AMSCs for transplantation purposes as well as for felines. We tested teratoma formation following cell injection into BALB/c nude mice and then assessed expression of haematopoietic, mesenchymal, tumorigenic, pluripotency and cellular regulation markers using flow cytometry and qPCR. The use of canine AMSCs did not result in macroscopic tumour formation as determined 60 days after transplantation. The immunophenotypic characterization by flow cytometry revealed expression of mesenchymal markers (CD73 and CD90) and expression of the pluripotent marker OCT4 and SOX2. Quantitative PCR analysis revealed that there were no differences in the patterns of gene expression (CD34, CD73, OCT4, CD30 and P53) between canine and feline AMSCs, with the exception of the expression of SOX2 and CD90.
多项研究探讨了再生医学中利用干细胞的创新程序的生物安全性。先前的研究表明，使用猫和人羊膜来源的干细胞（AMSCs）后未形成肿瘤。相比之下，使用犬类AMSCs时观察到了肿瘤形成。这些发现表明，猫和人的AMSCs适合用于细胞移植试验，而犬类AMSCs不适合。本研究旨在进一步评估将犬类AMSCs用于移植目的以及用于猫的可行性。我们将细胞注射到BALB/c裸鼠体内后测试畸胎瘤的形成，然后使用流式细胞术和定量聚合酶链反应（qPCR）评估造血、间充质、致瘤、多能性和细胞调节标志物的表达。移植60天后的检测结果显示，使用犬类AMSCs并未导致肉眼可见的肿瘤形成。流式细胞术的免疫表型特征分析显示间充质标志物（CD73和CD90）以及多能性标志物OCT4和SOX2的表达。定量PCR分析显示，除SOX2和CD90的表达外，犬类和猫类AMSCs之间的基因表达模式（CD34、CD73、OCT4、CD30和P53）没有差异。
