Generation of highly enriched V2a interneurons from mouse embryonic stem cells

Experimental Neurology
Nisha IyerShelly E Sakiyama-Elbert

Abstract

Challenges in parsing specific contributions to spinal microcircuit architecture have limited our ability to model and manipulate those networks for improved functional regeneration after injury or disease. While spinal interneurons (INs) have been implicated in driving coordinated locomotor behaviors, they constitute only a small percentage of the spinal cord and are difficult to isolate from primary tissue. In this study, we employed a genetic strategy to obtain large quantities of highly enriched mouse embryonic stem cell (ESC)-derived V2a INs, an excitatory glutamatergic IN population that is defined by expression of the homeodomain protein Chx10 during development. Puromycin N-acetyltransferase expression was driven by the native gene regulatory elements of Chx10 in the transgenic ESC line, resulting in positive selection of V2a INs after induction and treatment with puromycin. Directly after selection, approximately 80% of cells are Chx10(+), with 94% Lhx3(+); after several weeks, cultures remain free of proliferative cell types and mature into normal glutamatergic neurons as assessed by molecular markers and electrophysiological methods. Functional synapses were observed between selected ESC-derived V2a INs and motor neu...Continue Reading

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Citations

Aug 18, 2016·Biotechnology and Bioengineering·Nisha R IyerShelly E Sakiyama-Elbert
Dec 9, 2016·Expert Review of Neurotherapeutics·Michael A LaneItzhak Fischer
Aug 31, 2017·Stem Cells and Development·Russell E ThompsonShelly E Sakiyama-Elbert
Jun 7, 2018·Journal of Neurotrauma·Lyandysha V ZholudevaMichael A Lane
Oct 17, 2018·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Nicholas White, Shelly E Sakiyama-Elbert
Nov 2, 2018·Journal of Neuroscience Research·Lyandysha V Zholudeva, Michael A Lane
Jan 19, 2019·Cell Transplantation·Lyandysha V Zholudeva, Michael A Lane
Oct 20, 2019·Nature Protocols·Jessica C ButtsTodd C McDevitt
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Dec 13, 2017·Proceedings of the National Academy of Sciences of the United States of America·Kesavan MeganathanKristen L Kroll
May 16, 2018·Stem Cells International·E M SamoilovaV P Baklaushev
Feb 6, 2020·Frontiers in Systems Neuroscience·Victoria N JensenSteven A Crone
Jul 19, 2018·Trends in Neurosciences·Lyandysha V ZholudevaMichael A Lane
Apr 9, 2021·Expert Opinion on Biological Therapy·William Brett McIntyreMichael G Fehlings
May 25, 2021·Brain Research Bulletin·Aleeza Zilberman, R Chase Cornelison
Jun 19, 2021·Stem Cells and Development·Jennifer PardieckShelly Sakiyama-Elbert

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