Xu Lifa, Xu Congjing, Zhou Shuping, Liu Xueke, Wang Jian, Liu Xinkuang, Qian Suping, Xin Yingru, Gao Yi, Zhu Yongqiang, Tang Xiaolong
Stem Cell Engineering Research Center, School of Medical, Anhui University of Science & Technology Huainan 232001, P.R. China.
Department of Respiration, Tumour Hospital of Affiliated Huainan Oriental Hospital Group, Anhui University of Science and Technology Huainan 232035, P.R. China.
Am J Transl Res. 2017 Mar 15;9(3):874-886. eCollection 2017.
A shortage of postmortem pancreatic tissue for islet isolation impedes the application of cell replacement therapy in patients with diabetes. As an alternative for islet cell transplantation, transcription factors, including PDX1, PAX4, and neurogenin-3, that aid in the formation of insulin-producing β cells during development have been investigated. The present study evaluated the effects of PAX4 and PDX1 on the differentiation of mesenchymal stem cells (MSCs) into insulin-producing β-like cells using recombinant adenoviruses carrying PDX1 or PDX1 plus PAX4. RT-PCR, Western blot, and immunofluorescence assays were used to detect the expression levels of relevant genes and proteins, and enzyme-linked immunosorbent assays were used to determine the amount of insulin and C-peptide secreted by the virus-infected cells following stimulation with high glucose. The results showed that PAX4 markedly enhanced the propensity of PDX1-positive MSCs to form mature islet-like clusters and functional insulin-producing β-like cells. Our findings provide a novel foundation for generating β-like cells from MSCs with PAX4 and PDX1 for future clinical application.
用于胰岛分离的尸检胰腺组织短缺阻碍了细胞替代疗法在糖尿病患者中的应用。作为胰岛细胞移植的替代方法，人们研究了包括PDX1、PAX4和神经生成素-3在内的转录因子，这些转录因子在发育过程中有助于产生胰岛素的β细胞的形成。本研究使用携带PDX1或PDX1加PAX4的重组腺病毒评估了PAX4和PDX1对间充质干细胞（MSC）分化为产生胰岛素的β样细胞的影响。采用逆转录-聚合酶链反应（RT-PCR）、蛋白质印迹法和免疫荧光分析法检测相关基因和蛋白质的表达水平，并采用酶联免疫吸附测定法测定高糖刺激后病毒感染细胞分泌的胰岛素和C肽量。结果表明，PAX4显著增强了PDX1阳性MSC形成成熟胰岛样簇和功能性产生胰岛素的β样细胞的倾向。我们的研究结果为利用PAX4和PDX1从MSC生成β样细胞以供未来临床应用提供了新的基础。
