PCL/PVA nanofibrous scaffold improve insulin-producing cells generation from human induced pluripotent stem cells
PCL/PVA nanofibrous scaffold improve insulin-producing cells generation from human induced pluripotent stem cells
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DOI：
10.1016/j.gene.2018.05.115
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发表时间：
2018-09-10
期刊：
影响因子：
3.5
通讯作者：
Kehtari, Mousa
中科院分区：
文献类型：
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作者：
Abazari, Mohammad Foad;Soleimanifar, Fatemeh;Kehtari, Mousa
干细胞的胰腺分化将有助于I型糖尿病（T1 DM）患者的治疗。合成生物聚合物的利用提供了细胞外基质（ECM）和体外所需的属性，以增强干细胞增殖，附着和分化的条件。以聚己内酯和聚乙烯醇（PCL/PVA）为基质的混合支架，可建立体外三维（3D）培养模型。本研究的目的是研究人诱导多能干细胞（hiPSCs）在3D培养中向胰岛素产生细胞（IPC）分化的能力，并通过定量PCR和免疫荧光法分别在mRNA和蛋白水平上评估常规培养（2D）组与3D培养组的差异。通过葡萄糖刺激试验检测分化后IPC的C肽和胰岛素释放功能。Real-Time PCR结果显示iPSCs-IPC表达胰腺特异性转录因子（胰岛素、Pdxl、胰高血糖素、Glut 2和Ngn 3）。PCL/PVA支架中这些转录因子的表达均高于2D组。除IPC外，还通过免疫化学检测特异性标志物。两组中的这些细胞在通过ELISA的葡萄糖激发试验中分泌胰岛素和C肽，显示体外成熟。目前的研究结果表明，PCL/PVA纳米纤维支架可以增强IPC从hiPSCs的分化。本研究为β细胞替代治疗和胰腺组织工程治疗T1 DM提供了新的途径。
Pancreatic differentiation of stem cells will aid treatment of patients with type I diabetes mellitus (T1DM). Synthetic biopolymers utilization provided extracellular matrix (ECM) and desired attributes in vitro to enhance conditions for stem cells proliferation, attachment and differentiation. A mixture of polycaprolactone and polyvinyl alcohol (PCL/PVA)-based scaffold, could establish an in vitro three-dimensional (3D) culture model. The objective of this study was investigation of the human induced pluripotent stem cells (hiPSCs) differentiation capacity to insulin-producing cells (IPCs) in 3D culture were compared with conventional culture (2D) groups evaluated at the mRNA and protein levels by quantitative PCR and immunofluorescence assay, respectively. The functionality of differentiated IPCs was assessed by C-peptide and insulin release in response to glucose stimulation test. Real-Time PCR results showed that iPSCs-IPCs expressed pancreas-specific transcription factors (Insulin, Pdxl, Glucagon, Glut2 and Ngn3). The expressions of these transcription factors in PCL/PVA scaffold were higher than 2D groups. In addition to IPCs specific markers were detected by immunochemistry. These cells in both groups secreted insulin and C-peptide in a glucose challenge test by ELISA showing in vitro maturation. The results of current study demonstrated that enhanced differentiation of IPCs from hiPSCs could be result of PCL/PVA nanofibrous scaffolds. In conclusion, this research could provide a new approach to beta like cells replacement therapies and pancreatic tissue engineering for T1DM in the future.