AUTHOR=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 TITLE=Different iPSC-derived neural stem cells shows various spectrums of spontaneous differentiation during long term cultivation JOURNAL=Frontiers in Molecular Neuroscience VOLUME=16 YEAR=2023 URL=https://www.frontiersin.org/journals/molecular-neuroscience/articles/10.3389/fnmol.2023.1037902 DOI=10.3389/fnmol.2023.1037902 ISSN=1662-5099 ABSTRACT=Introduction

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.

Methods

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.

Results

We found that various NSC lines generate significantly different spectrums of differentiated neural cells, which can also change significantly during long-term cultivation in vitro.

Discussion

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 in vitro.