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首页> 美国卫生研究院文献>Journal of Clinical Research in Pediatric Endocrinology >Generation of Monocyte-Derived Insulin-Producing Cells from Non-Human Primates According to an Optimized Protocol for the Generation ofPCMO-Derived Insulin-Producing Cells
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Generation of Monocyte-Derived Insulin-Producing Cells from Non-Human Primates According to an Optimized Protocol for the Generation ofPCMO-Derived Insulin-Producing Cells

机译：根据用于生成PCMO的胰岛素产生细胞的优化方案从非人类灵长类动物产生单核细胞的产生胰岛素的细胞

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>Ob­jec­ti­ve: The vision of potential autologous cell therapy for the cure of diabetes encourages ongoing research. According to a previously published protocol for the generation of insulin-producing cells from human monocytes, we analyzed whether the addition of growth factors could increase insulin production. This protocol was then transferred to a non-human primate model by using either blood- or spleen-derived monocytes. >Methods: Human monocytes were treated to dedifferentiate into programmable cells of monocytic origin (PCMO). In addition to the published protocol, PCMOs were then treated with either activin A, betacellulin, exendin 3 or 4. Cells were characterized by protein expression of insulin, Pdx-1, C-peptide and Glut-2. After identifying the optimal protocol, monocytes from baboon blood were isolated and the procedure was repeated. Spleen monocytes following splenectomy of a live baboon were differentiated and analyzed in the same manner and calculated in number and volume.>Results: Insulin content of human cells was highest when cells were treated with activin A and their insulin content was 13 000 µU/1 million cells. Insulin-producing cells form primate monocytes could successfully be generated despite using human growth factors and serum. Expression of insulin, Pdx-1, C-peptide and Glut-2 was comparable to that of human neo-islets. Total insulin content of activin A-treated baboon monocytes was 16 000 µU/1 million cells.>Conclusion: We were able to show that insulin-producing cells can be generated from baboon monocytes with human growth factors. The amount generated from one spleen could be enough to cure a baboon from experimentally induced diabetes in an autologous cell transplant setting.
机译：>目标：对潜在的自体细胞疗法治疗糖尿病的愿景鼓励了正在进行的研究。根据先前发布的从人单核细胞生成胰岛素产生细胞的协议，我们分析了添加生长因子是否可以增加胰岛素产生。然后通过使用血液或脾脏衍生的单核细胞将该方案转移到非人灵长类动物模型中。 >方法：将人类单核细胞去分化为单核细胞来源的可编程细胞（PCMO）。除已公开的方案外，然后用激活素A，β-纤维素，exendin 3或4处理PCMO。细胞以胰岛素，Pdx-1，C-肽和Glut-2的蛋白表达为特征。确定最佳方案后，从狒狒血液中分离出单核细胞，并重复该过程。用同样的方法对脾狒狒脾切除后的脾单核细胞进行分化和分析，并计算其数量和体积。>结果：用激活素A处理细胞及其胰岛素含量后，人体细胞的胰岛素含量最高是13000 µU / 1百万个细胞。尽管使用了人类生长因子和血清，但仍可以成功生成灵长类单核细胞形成的胰岛素产生细胞。胰岛素，Pdx-1，C肽和Glut-2的表达与人新胰岛相当。激活素A处理过的狒狒单核细胞的总胰岛素含量为16000 µU / 1百万个细胞。>结论：我们能够证明，具有人类生长因子的狒狒单核细胞可以产生胰岛素生产细胞。在一个自体细胞移植的环境中，一个脾脏产生的数量足以治愈一只实验性糖尿病引起的狒狒。

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