Generation of GABAergic striatal neurons by a novel iPSC differentiation protocol enabling scalability and cryopreservation of progenitor cells.

Cytotechnology
Elena V Grigor'evaAnastasia A Malakhova

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 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...Continue Reading

References

Dec 5, 1995·Proceedings of the National Academy of Sciences of the United States of America·F H GageJ Ray
Sep 6, 1996·Science·B J KnowltonL R Squire
Apr 30, 2004·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Zachary C BaquetKevin R Jones
Aug 18, 2004·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Miguel A López-ToledanoEulalia Bazán
Feb 3, 2005·Nature Biotechnology·Xue-Jun LiSu-Chun Zhang
Nov 8, 2005·Current Opinion in Neurobiology·Ann M Graybiel
Mar 24, 2007·Progress in Neurobiology·Chiara Zuccato, Elena Cattaneo
Oct 17, 2008·Proceedings of the National Academy of Sciences of the United States of America·Laetitia AubryAnselme L Perrier
Jun 3, 2009·Stem Cells·Andrea Abdipranoto-CowleyBryce Vissel
Oct 8, 2009·Journal of Drug Targeting·Sandra R Alcalá-BarrazaLinda K McLoon
Jan 29, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Nao ChuhmaStephen Rayport
Apr 5, 2011·Nature Methods·Keisuke OkitaShinya Yamanaka
Jul 26, 2011·Cellular and Molecular Life Sciences : CMLS·Huisheng Liu, Su-Chun Zhang
Oct 16, 2015·Biochemistry Insights·Han WangJake Leighton
Apr 16, 2016·Molecular Neurodegeneration·Evgeny D NekrasovSergey L Kiselev

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adult Stem Cells

Adult stem cells reside in unique niches that provide vital cues for their survival, self-renewal, and differentiation. They hold great promise for use in tissue repair and regeneration as a novel therapeutic strategies. Here is the latest research.