Kashfi Shirin, Peymani Maryam, Ghaedi Kamran, Baharvand Hossein, Nasr-Esfahani Mohammad Hossein, Javan Mohammad
Department of Developmental Biology, University of Science and Culture, Tehran, Iran.
Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Cell J. 2017 Oct;19(3):386-402. doi: 10.22074/cellj.2017.3906. Epub 2017 Aug 19.
Due to recent progress in production of human embryonic stem cell-derived oligodendrocyte progenitor cells (hESC-OPCs) for ameliorating myelin disease such as multiple sclerosis (MS) and the role of purinergic signaling in OPCs development, we avaluated the profile of purinergic receptors expression during development of OPCs from hESC.
In this experimental study, we used reverse transcription and quantitative polymerase chain reaction (RT-qPCR) to obtain more information about potential roles of purinergic receptors during in vitro production of hESC-OPCs. We first determined the expression level of different subtypes of purinergic receptors in hESCs, embryoid bodies (EBs), and hESC-OPCs. The effects of A1 adenosine receptor (A1AR) activation on hESC-OPCs development were subsequently examined.
hESCs and OPCs had different mRNA expression levels of the AR subtypes. ARs mRNA were expressed in the EB stage, except for A2AAR. We observed expressions of several P2X (P2X1, 2, 3, 4, 5, 7) and P2Y (P2Y1, 2, 4, 6, 11-14) genes in hESCs. hESC-OPCs expressed different subtypes of P2X (P2X1, 2, 3,4,5,7) and P2Y (P2Y1, 2, 4, 6, 11-14). Except for P2X1 and P2X6, all other P2X and P2Y purinergic receptor subtypes expressed in EBs. We also indicate that A1AR might be involved in modulating gene expression levels of cell cycle regulators in an agonist and/or dose-dependent manner.
Elucidation of the expression pattern of purinergic receptors and the effects of different subtypes of these receptors in hESC-OPCs may have a promising role in future cell-based therapy or drug design for demyelinating disease.
鉴于在生产用于改善髓鞘疾病（如多发性硬化症，MS）的人胚胎干细胞来源的少突胶质前体细胞（hESC-OPCs）方面取得的最新进展以及嘌呤能信号在OPCs发育中的作用，我们评估了hESC来源的OPCs发育过程中嘌呤能受体的表达情况。
在本实验研究中，我们使用逆转录和定量聚合酶链反应（RT-qPCR）来获取更多关于嘌呤能受体在hESC-OPCs体外生产过程中潜在作用的信息。我们首先测定了嘌呤能受体不同亚型在hESCs、胚状体（EBs）和hESC-OPCs中的表达水平。随后研究了A1腺苷受体（A1AR）激活对hESC-OPCs发育的影响。
hESCs和OPCs中AR亚型的mRNA表达水平不同。除A2AAR外，ARs mRNA在EB阶段表达。我们在hESCs中观察到几种P2X（P2X1、2、3、4、5、7）和P2Y（P2Y1、2、4、6、11 - 14）基因的表达。hESC-OPCs表达不同亚型的P2X（P2X1、2、3、4、5、7）和P2Y（P2Y1、2、4、6、11 - 14）。除P2X1和P2X6外，所有其他P2X和P2Y嘌呤能受体亚型在EBs中表达。我们还指出，A1AR可能以激动剂和/或剂量依赖的方式参与调节细胞周期调节因子的基因表达水平。
阐明嘌呤能受体的表达模式以及这些受体不同亚型在hESC-OPCs中的作用，可能在未来基于细胞治疗或脱髓鞘疾病药物设计中发挥重要作用。
