Insulin Producing Cells from Amniotic Stem Cells for Diabetes Therapy

糖尿病是一个日益严重的世界性问题。在美国，它影响了1800多万人，并导致 每年的医疗保健费用超过1300亿美元。1型糖尿病的胰岛素治疗，以及在晚期糖尿病 2型糖尿病，不能预防严重的长期并发症，包括神经病变，血管疾病， 视网膜病变和肾功能衰竭。胰岛移植以恢复胰岛素的产生具有重要意义 我保证。然而，供体胰腺的供应远远不能满足医疗需求。新的资源来源 需要产生胰岛素的细胞才能充分发挥糖尿病细胞疗法的潜力。我们建议 从羊水中分离的干细胞通过体外分化产生胰岛β系细胞。这些 “AFS细胞”既能广泛扩增，又能分化为所有三个胚胎的衍生细胞。 胚芽层。我们的初步研究表明，小鼠AFS细胞可以产生胰岛素产生细胞和胰岛- 胰腺转录因子PDX-1的表达促进了类细胞团(“新胰岛”)的形成。我们现在 建议从人和非人灵长类(NHP)AFS细胞中培养新的胰岛。要高效地生成 在胰岛素分泌细胞中，我们将优化输送一个表达PDX-1的质粒载体，并将系统地 测试生长因子和之前显示的促进胰岛β细胞分化的底物。这个 所产生的新胰岛将与真实的人类和NHP胰岛进行比较，使用为 临床移植。评估他们恢复对葡萄糖代谢的控制和产生 胰岛素和C肽，新的胰岛将被植入链脲佐菌素糖尿病免疫缺陷小鼠体内 (STZ)。STZ治疗的NHP中的移植将在生理学模型中评估新生胰岛的功能 更接近于人类。NHP AFS细胞系将在孕妇羊膜穿刺术后获得。这些 干细胞将被用来产生新的胰岛，用于自体移植到相应的后代中。 同样的供体细胞将在使用临床相关免疫抑制的异基因受者中进行比较。 养生法。成功开发出来源丰富的可移植胰岛素产生细胞 将对一个重大公共卫生问题的治疗产生深远影响。

Diabetes mellitus is a growing problem worldwide. In the US it affects over 18 million people and results in annual health care costs exceeding $130 billion. Insulin therapy of Type 1 diabetes, and in advanced cases of Type 2 diabetes, does not prevent serious long-term complications including neuropathy, vascular disease, retinopathy and renal failure. Transplantation of pancreatic islets to restore insulin production offers significant promise. However, the supply of donor pancreata falls far short of meeting the medical need. New sources of insulin producing cells will be required to realize the full potential of cell therapy for diabetes. We propose to generate pancreatic beta lineage cells by in vitro differentiation of stem cells isolated from amniotic fluid. These "AFS cells" are capable of both extensive expansion and differentiation into derivatives of all three embryonic germ layers. Our Preliminary Studies showed that mouse AFS cells can yield insulin producing cells and islet- like cell clusters ("neo-islets"), promoted by expression of the pancreatic transcription factor PDX-1. We now propose to produce neo-islets from human and nonhuman primate (NHP) AFS cells. To efficiently generate insulin producing cells, we will optimize delivery of a plasmid vector to express PDX-1, and will systematically test growth factors and substrates shown previously to promote pancreatic beta cell differentiation. The resulting neo-islets will be compared with authentic human and NHP pancreatic islets using tests developed for clinical transplantation. To assess their ability to restore control of glucose metabolism and production of insulin and C-peptide, neo-islets will be implanted in immunodeficient mice made diabetic with streptozotocin (STZ). Transplantation in STZ-treated NHP will assess the function of neo-islets in a model physiologically more similar to humans. NHP AFS cell lines will be derived after amniocentesis of pregnant mothers. These stem cells will be used to generate neo-islets for autologous transplantation into the corresponding offspring. The same donor cells will be compared in allogeneic recipients using clinically relevant immunosuppression regimens. Successful development of an abundant source of transplantable insulin producing cells potentially would have a profound impact on the treatment of a major public health problem.