Galiakberova Adelya Albertovna, Brovkina Olga Igorevna, Kondratyev Nikolay Vitalyevich, Artyuhov Alexander Sergeevich, Momotyuk Ekaterina Dmitrievna, Kulmukhametova Olga Nikolaevna, Lagunin Alexey Aleksandrovich, Shilov Boris Vladimirovich, Zadorozhny Anton Dmitrievich, Zakharov Igor Sergeevitch, Okorokova Larisa Sergeevna, Golimbet Vera Evgenievna, Dashinimaev Erdem Bairovich
Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, Moscow, Russia.
Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia.
Front Mol Neurosci. 2023 May 2;16:1037902. doi: 10.3389/fnmol.2023.1037902. eCollection 2023.
Culturing of human neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSC) is a promising area of research, as these cells have the potential to treat a wide range of neurological, neurodegenerative and psychiatric diseases. However, the development of optimal protocols for the production and long-term culturing of NSCs remains a challenge. One of the most important aspects of this problem is to determine the stability of NSCs during long-term in vitro passaging. To address this problem, our study was aimed at investigating the spontaneous differentiation profile in different iPSC-derived human NSCs cultures during long-term cultivation using.
Four different IPSC lines were used to generate NSC and spontaneously differentiated neural cultures using DUAL SMAD inhibition. These cells were analyzed at different passages using immunocytochemistry, qPCR, bulk transcriptomes and scRNA-seq.
We found that various NSC lines generate significantly different spectrums of differentiated neural cells, which can also change significantly during long-term cultivation .
Our results indicate that both internal (genetic and epigenetic) and external (conditions and duration of cultivation) factors influence the stability of NSCs. These results have important implications for the development of optimal NSCs culturing protocols and highlight the need to further investigate the factors influencing the stability of these cells .
源自诱导多能干细胞（iPSC）的人类神经干细胞（NSC）培养是一个很有前景的研究领域，因为这些细胞有潜力治疗多种神经、神经退行性和精神疾病。然而，开发用于NSC生产和长期培养的最佳方案仍然是一项挑战。这个问题最重要的方面之一是确定NSC在长期体外传代过程中的稳定性。为了解决这个问题，我们的研究旨在调查不同iPSC来源的人类NSC培养物在长期培养过程中的自发分化情况。
使用四种不同的iPSC系通过双SMAD抑制来生成NSC和自发分化的神经培养物。使用免疫细胞化学、qPCR、批量转录组和scRNA-seq对这些细胞在不同传代时进行分析。
我们发现不同的NSC系产生明显不同的分化神经细胞谱，并且在长期培养过程中也会发生显著变化。
我们的结果表明，内部（遗传和表观遗传）和外部（培养条件和持续时间）因素都会影响NSC的稳定性。这些结果对开发最佳的NSC培养方案具有重要意义，并突出了进一步研究影响这些细胞稳定性因素的必要性。
