Comparison of two types of non-adherent plate for neuronal differentiation of mouse embryonic stem cells
Comparison of two types of non-adherent plate for neuronal differentiation of mouse embryonic stem cells
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两种非贴壁板对小鼠胚胎干细胞神经元分化的比较
DOI：
10.1007/s10616-016-9968-z
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发表时间：
2016
期刊：
影响因子：
2.2
通讯作者：
Kiyosawa H
中科院分区：
文献类型：
--
作者：
Eitoku M;Suganuma N;Kiyosawa H
小鼠胚胎干细胞（ESCs）体外分化系统被广泛用于细胞分化、器官发生和再生医学的研究。我们利用F1杂交ESCs的体外神经元分化研究了神经元特异性印迹基因Ube3a及其反义转录物（Ube3a ATS）的调控。在分化过程中，用于胚状体（EB）形成的每一种不同的非贴壁板都有不同的成本；值得注意的是，涂有2-甲基丙烯酰氧乙基磷酸胆碱（MPC）聚合物的板比未经处理的聚苯乙烯板更昂贵。在这里，我们评估了聚合物涂布板是否比未处理的板提供更好的结果。分化的第一阶段是在MPC聚合物涂层或未经处理的板上进行的。然后将形成的EBs传代到层粘连蛋白涂层板上进一步分化为神经元。神经元特异性印迹状态和神经元特异性标记物sube3a atsandmtap2的表达水平在未处理的和MPC聚合物包被的神经元上均无差异。上述结果表明，两种非贴壁板在诱导小鼠ESCs神经元分化和EB形成方面表现出几乎相同的特性。我们的研究证明，未经处理的聚苯乙烯板是小鼠ESCs体外分化系统中EB形成的一种经济有效的选择。
In vitro differentiation systems of mouse embryonic stem cells (ESCs) are widely used as tools for studies of cell differentiation, organogenesis, and regenerative medicine. We have studied the regulation of neuron-specific imprinting genes,Ube3aand its antisense transcripts (Ube3a ATS), using in vitro neuronal differentiation of F1 hybrid ESCs. Each different non-adherent plate used for embryoid body (EB) formation during differentiation is associated with different costs; notably, plates coated with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer are more expensive than untreated polystyrene plates. Here, we assessed whether the polymer-coated plates gave better results than the untreated plates. The first stage of differentation was performed in the MPC polymer-coated or untreated plates. The formed EBs were then passaged onto laminin-coated plates for further differentiation into neurons. Neither the neuron-specific imprinting status ofUbe3anor the expression levels of the neuron-specific markersUbe3a ATSandMtap2differed between neurons prepared on untreated plates and those prepared on MPC polymer-coated plates. These results suggest that the two non-adherent plates displayed almost the same characteristics for inducing neuronal differentiation of mouse ESCs and EB formation. Our study proved that untreated polystyrene plates are a cost-effective choice for EB formation in in vitro differentiation systems of mouse ESCs.