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

Stem Cell Research & Therapy

Ling Wei³  Dongdong Chen³  Xiaohuan Gu³  Ying Guo¹  Osama Mohamad³  Mingke Song³  Danielle Drury-Stewart² 

[1] Department of Biostatistics and Bioinformatics, Emory University, 1518 Clifton Road, NE, Atlanta, GA 30322, USA;Department of Biomedical Engineering, Georgia Institute of Technology, 15 Ferst Drive, NW, Atlanta, GA 30332, USA;Department of Anesthesiology, Emory University, 101 Woodruff Circle, Atlanta, GA 30322, USA

关键词: Small molecule;    Neurogenesis;    Ischemic stroke;    Cell therapy;    Stem cell;    Electrophysiology;    Neural precursor;    Human embryonic stem cell;   

Others  :  847125
DOI  :  10.1186/scrt292

 received in 2013-01-11, accepted in 2013-07-26,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

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

【 图 表 】

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