当前位置: X-MOL 学术Cytotechnology › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Generation of GABAergic striatal neurons by a novel iPSC differentiation protocol enabling scalability and cryopreservation of progenitor cells.
Cytotechnology ( IF 2.2 ) Pub Date : 2020-06-09 , DOI: 10.1007/s10616-020-00406-7
Elena V Grigor'eva 1 , Tuyana B Malankhanova 1 , Aizhan Surumbayeva 1 , Sophia V Pavlova 1 , Julia M Minina 1 , Elena A Kizilova 1 , Lyubov A Suldina 1 , Ksenia N Morozova 1 , Elena Kiseleva 1 , Eugeny D Sorokoumov 1 , Igor N Lebedev 2 , Suren M Zakian 1 , Anastasia A Malakhova 1
Affiliation  

Cell models are promising tools for studying hereditary human neurodegenerative diseases. Neuronal derivatives of pluripotent stem cells provide the opportunity to investigate different stages of the neurodegeneration process. Therefore, easy and large-scale production of relevant cell types is a crucial barrier to overcome. In this work, we present an alternative protocol for iPSC differentiation into GABAergic medium spiny neurons (MSNs). The first stage involved dual-SMAD signalling inhibition through treatment with SB431542 and LDN193189, which results in the generation of neuroectodermal cells. Moreover, we used bFGF as a neuronal survival factor and dorsomorphin to inhibit BMP signalling. The combined treatment of dorsomorphin and SB431542 significantly enhanced neuronal induction, which was confirmed by the increased expression of the telencephalic-specific markers SOX1 and OTX2 as well as the forebrain marker PAX6. The next stage involved the derivation of actively proliferating MSN progenitor cells. An important feature of our protocol at this stage is the ability to perform prolonged cultivation of precursor cells at a high density without losing phenotypic properties. Moreover, the protocol enables multiple expansion steps (> 180 days cultivation) and cryopreservation of MSN progenitors. Therefore, this method allows quick production of a large number of neurons that are relevant for basic research, large-scale drug screening, and toxicological studies.



中文翻译:

通过新型 iPSC 分化协议生成 GABA 能纹状体神经元,从而实现祖细胞的可扩展性和冷冻保存。

细胞模型是研究遗传性人类神经退行性疾病的有前途的工具。多能干细胞的神经元衍生物为研究神经变性过程的不同阶段提供了机会。因此,相关细胞类型的简单和大规模生产是需要克服的关键障碍。在这项工作中,我们提出了一种将 iPSC 分化为 GABA 能介质多刺神经元 (MSN) 的替代方案。第一阶段涉及通过 SB431542 和 LDN193189 处理的双重 SMAD 信号抑制,这导致神经外胚层细胞的产生。此外,我们使用 bFGF 作为神经元存活因子和 dorsomorphin 来抑制 BMP 信号传导。dorsomorphin 和 SB431542 的联合治疗显着增强了神经元诱导,SOX1OTX2以及前脑标记PAX6。下一阶段涉及积极增殖的 MSN 祖细胞的衍生。在这个阶段,我们的协议的一个重要特点是能够在不失去表型特性的情况下以高密度长时间培养前体细胞。此外,该协议支持多个扩展步骤(> 180 天培养)和 MSN 祖细胞的冷冻保存。因此,这种方法可以快速生产大量与基础研究、大规模药物筛选和毒理学研究相关的神经元。

更新日期:2020-06-09
down
wechat
bug