Boroujeni Zahra Niki, Aleyasin Ahmad
National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Biotechnol Appl Biochem. 2014 Mar-Apr;61(2):82-92. doi: 10.1002/bab.1127. Epub 2014 Mar 20.
Diabetes mellitus is characterized by autoimmune destruction of pancreatic beta cells, leading to decreased insulin production. Differentiation of mesenchymal stem cells (MSCs) into insulin-producing cells offers novel ways of diabetes treatment. MSCs can be isolated from the human umbilical cord tissue and differentiate into insulin-secreting cells. Human umbilical cord-derived stem cells (hUDSCs) were obtained after birth, selected by plastic adhesion, and characterized by flow cytometric analysis. hUDSCs were transduced with nonintegrated lentivirus harboring PDX1 (nonintegrated LV-PDX1) and was cultured in differentiation medium in 21 days. Pancreatic duodenum homeobox protein-1 (PDX1) is a transcription factor in pancreatic development. Significant expressions of PDX1, neurogenin3 (Ngn3), glucagon, glucose transporter2 (Glut2), and somatostatin were detected by quantitative RT-PCR (P < 0.05). PDX1 and insulin proteins were shown by immunocytochemistry analysis. Insulin secretion of hUDSCs(PDX1+) in the high-glucose medium was 1.8 μU/mL. They were used for treatment of diabetic rats and could decrease the blood glucose level from 400 mg/dL to a normal level in 4 days. In conclusion, our results demonstrated that hUDSCs are able to differentiate into insulin-producing cells by transduction with nonintegrated LV-PDX1. These hUDSCs(PDX1+) have the potential to be used as a viable resource in cell-based gene therapy of type 1 diabetes.
糖尿病的特征是胰腺β细胞发生自身免疫性破坏，导致胰岛素分泌减少。间充质干细胞（MSCs）向胰岛素分泌细胞的分化为糖尿病治疗提供了新途径。MSCs可从人脐带组织中分离出来，并分化为胰岛素分泌细胞。人脐带来源的干细胞（hUDSCs）在出生后获得，通过塑料贴壁法进行筛选，并通过流式细胞术分析进行鉴定。用携带PDX1的非整合慢病毒（非整合LV-PDX1）转导hUDSCs，并在分化培养基中培养21天。胰腺十二指肠同源盒蛋白-1（PDX1）是胰腺发育中的一种转录因子。通过定量RT-PCR检测到PDX1、神经源性分化因子3（Ngn3）、胰高血糖素、葡萄糖转运蛋白2（Glut2）和生长抑素的显著表达（P < 0.05）。通过免疫细胞化学分析显示了PDX1和胰岛素蛋白。hUDSCs（PDX1+）在高糖培养基中的胰岛素分泌量为1.8 μU/mL。它们被用于治疗糖尿病大鼠，并能在4天内将血糖水平从400 mg/dL降至正常水平。总之，我们的结果表明，hUDSCs通过用非整合LV-PDX1转导能够分化为胰岛素分泌细胞。这些hUDSCs（PDX1+）有潜力作为1型糖尿病基于细胞的基因治疗中的一种可行资源。
