Generation of GABAergic striatal neurons by a novel iPSC differentiation protocol enabling scalability and cryopreservat
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ORIGINAL ARTICLE
Generation of GABAergic striatal neurons by a novel iPSC differentiation protocol enabling scalability and cryopreservation of progenitor cells Elena V. Grigor’eva . Tuyana B. Malankhanova . Aizhan Surumbayeva . Sophia V. Pavlova . Julia M. Minina . Elena A. Kizilova . Lyubov A. Suldina . Ksenia N. Morozova . Elena Kiseleva . Eugeny D. Sorokoumov . Igor N. Lebedev . Suren M. Zakian . Anastasia A. Malakhova Received: 15 October 2019 / Accepted: 31 May 2020 Ó Springer Nature B.V. 2020
Abstract 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 largescale 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 Elena V. Grigor’eva and Tuyana B. Malankhanova contributed equally to this work.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10616-020-00406-7) contains supplementary material, which is available to authorized users. E. V. Grigor’eva (&) T. B. Malankhanova A. Surumbayeva S. V. Pavlova J. M. Minina E. A. Kizilova L. A. Suldina K. N. Morozova E. Kiseleva E. D. Sorokoumov S. M. Zakian A. A. Malakhova Federal Research Center Institute of Cytology and Genetics, The Siberian Branch of the Russian Academy of Sciences, 10 Lavrentiev Ave, Novosibirsk, Russian Federation 630090 e-mail: [email protected] T. B. Malankhanova e-mail: [email protected] A. Surumbayeva e-mail: [email protected]
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 S. V. Pavlova e-mail: [email protected] J. M. Minina e-mail: [email protected] E. A. Kizilova e-mail: [email protected] L. A. Suldina e-mail: [email protected] K. N. Morozova e-mail: [email protected] E. Kiseleva e-mail: [email protected]
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Cytotechnology
large number of neurons that are relevant for basic research, large-scale d
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