Department of Nanobiotechnology, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran.
Mol Biol Rep. 2014 Mar;41(3):1713-21. doi: 10.1007/s11033-014-3020-1. Epub 2014 Jan 29.
Although embryonic stem cells (ESCs) have enormous potentials due to their pluripotency, their therapeutic use is limited by ethical, biological and safety issues. Compared to ESCs, induced pluripotent stem cells (iPSCs) can be obtained from mouse or human fibroblasts by reprogramming. Numerous studies have established many protocols for differentiation of human iPSCs (hiPSCs) into neural lineages. However, the low differentiation efficiency of such protocols motivates researchers to design new protocols for high yield differentiation. Herein, we compared neural differentiation potential of three induction media for conversion of hiPSCs into neural lineages. In this study, hiPSCs-derived embryoid bodies were plated on laminin coated dishes and were treated with three induction media including (1) bFGF, EGF (2) RA and (3) forskolin, IBMX. Immunofluorescence staining and quantitative real-time PCR (qPCR) analysis were used to detect the expression of neural genes and proteins. qPCR analysis showed that the expression of neural genes in differentiated hiPSCs in forskolin, IBMX supplemented media was significantly higher than undifferentiated cells and those in induction media containing bFGF, EGF or RA. In conclusion, our results indicated a successful establishment protocol with high efficiency for differentiation of hiPSCs into neural lineages.
虽然胚胎干细胞（ESCs）具有多能性，因此具有巨大的潜力，但它们的治疗用途受到伦理、生物和安全问题的限制。与 ESCs 相比，诱导多能干细胞（iPSCs）可以通过重编程从小鼠或人成纤维细胞中获得。许多研究已经建立了许多将人 iPSCs（hiPSCs）分化为神经谱系的方案。然而，这些方案的低分化效率促使研究人员设计新的方案以实现高产量分化。在此，我们比较了三种诱导培养基在将 hiPSCs 转化为神经谱系中的神经分化潜力。在这项研究中，hiPSCs 衍生的类胚体被接种在层粘连蛋白包被的培养皿上，并分别用三种诱导培养基处理，包括（1）bFGF、EGF，（2）RA 和（3）forskolin、IBMX。免疫荧光染色和实时定量 PCR（qPCR）分析用于检测神经基因和蛋白质的表达。qPCR 分析显示，在 forskolin、IBMX 补充培养基中分化的 hiPSCs 中神经基因的表达明显高于未分化细胞以及含有 bFGF、EGF 或 RA 的诱导培养基中的细胞。总之，我们的结果表明建立了一种高效的 hiPSCs 向神经谱系分化的方案。
