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Author Notes:

Corresponding author: Ling Wei, Email: lwei7@emory.edu.

DD-S carried out the cell and tissue processing, staining, Western blots, and most statistical analyses; participated in the study design; and drafted the manuscript.

MS carried out the electrophysiological measurements.

OM aided in cell culture, study design, and manuscript revision.

YG designed and carried out the statistical analysis of the adhesive removal tests and contributed to the drafting of the manuscript.

XG carried out all animal surgeries.

DC aided in immunohistochemistry and cell quantification.

LW helped in initiation of the research idea, participated in the study design, and revised the manuscript.

All authors read and approved the final manuscript.

The authors are grateful for helpful discussion with Shan Ping Yu and his support during this investigation.

The authors declare that they have no competing interests.

Subjects:

Research Funding:

This work was supported by National Institutes of Health (NIH) grants NS0458710, NS 057255, NS075338, and NS062097.

This work was also supported by the NIH grant C06 RR015455 from the Extramural Research Facilities Program of the National Center for Research Resources.

Keywords:

  • Human embryonic stem cell
  • Neural precursor
  • Electrophysiology
  • Stem cell
  • Cell therapy
  • Ischemic stroke
  • Neurogenesis
  • Small molecule

Highly efficient differentiation of neural precursors from human embryonic stem cells and benefits of transplantation after ischemic stroke in mice

Journal Title:

Stem Cell Research & Therapy

Volume:

Volume 4, Number 4

Publisher:

Type of Work:

Article | Final Publisher PDF

Abstract:

Introduction: Ischemic stroke is a leading cause of death and disability, but treatment options are severely limited. Cell therapy offers an attractive strategy for regenerating lost tissues and enhancing the endogenous healing process. In this study, we investigated the use of human embryonic stem cell-derived neural precursors as a cell therapy in a murine stroke model. Methods: Neural precursors were derived from human embryonic stem cells by using a fully adherent SMAD inhibition protocol employing small molecules. The efficiency of neural induction and the ability of these cells to further differentiate into neurons were assessed by using immunocytochemistry. Whole-cell patch-clamp recording was used to demonstrate the electrophysiological activity of human embryonic stem cell-derived neurons. Neural precursors were transplanted into the core and penumbra regions of a focal ischemic stroke in the barrel cortex of mice. Animals received injections of bromodeoxyuridine to track regeneration. Neural differentiation of the transplanted cells and regenerative markers were measured by using immunohistochemistry. The adhesive removal test was used to determine functional improvement after stroke and intervention. Results: After 11 days of neural induction by using the small-molecule protocol, over 95% of human embryonic stem-derived cells expressed at least one neural marker. Further in vitro differentiation yielded cells that stained for mature neuronal markers and exhibited high-amplitude, repetitive action potentials in response to depolarization. Neuronal differentiation also occurred after transplantation into the ischemic cortex. A greater level of bromodeoxyuridine co-localization with neurons was observed in the penumbra region of animals receiving cell transplantation. Transplantation also improved sensory recovery in transplant animals over that in control animals. Conclusions: Human embryonic stem cell-derived neural precursors derived by using a highly efficient small-molecule SMAD inhibition protocol can differentiate into electrophysiologically functional neurons in vitro. These cells also differentiate into neurons in vivo, enhance regenerative activities, and improve sensory recovery after ischemic stroke.

Copyright information:

© 2013 Drury-Stewart et al.; licensee BioMed Central Ltd.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 2.0 Generic License (http://creativecommons.org/licenses/by/2.0/).

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