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Inhibitor of PI3K/Akt Signaling Pathway Small Molecule Promotes Motor Neuron Differentiation of Human Endometrial Stem Cells Cultured on Electrospun Biocomposite Polycaprolactone/Collagen Scaffolds

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Abstract

Small molecules as useful chemical tools can affect cell differentiation and even change cell fate. It is demonstrated that LY294002, a small molecule inhibitor of phosphatidylinositol 3-kinase (PI3K)/Akt signal pathway, can inhibit proliferation and promote neuronal differentiation of mesenchymal stem cells (MSCs). The purpose of this study was to investigate the differentiation effect of Ly294002 small molecule on the human endometrial stem cells (hEnSCs) into motor neuron-like cells on polycaprolactone (PCL)/collagen scaffolds. hEnSCs were cultured in a neurogenic inductive medium containing 1 μM LY294002 on the surface of PCL/collagen electrospun fibrous scaffolds. Cell attachment and viability of cells on scaffolds were characterized by scanning electron microscope (SEM) and 3-(4,5-dimethylthiazoyl-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay. The expression of neuron-specific markers was assayed by real-time PCR and immunocytochemistry analysis after 15 days post induction. Results showed that attachment and differentiation of hEnSCs into motor neuron-like cells on the scaffolds with Ly294002 small molecule were higher than that of the cells on tissue culture plates as control group. In conclusion, PCL/collagen electrospun scaffolds with Ly294002 have potential for being used in neural tissue engineering because of its bioactive and three-dimensional structure which enhances viability and differentiation of hEnSCs into neurons through inhibition of the PI3K/Akt pathway. Thus, manipulation of this pathway by small molecules can enhance neural differentiation.

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Acknowledgments

The authors thank Tehran University of Medical Sciences to support this research with grant number “93-04-159-28027”.

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Authors and Affiliations

  1. Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Somayeh Ebrahimi-Barough, Jafar Ai & Roksana Tajerian

  2. Department of Biology, Faculty of Sciences, Shahid Chamran University , Ahvaz, Iran

    Elham Hoveizi

  3. Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Meysam Yazdankhah

  4. Division of Biomedical Engineering, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, North Karegar Ave., P.O. Box 14395-1561, Tehran, Iran

    Mehrdad Khakbiz

  5. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran

    Faezeh Faghihi

  6. Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran

    Neda Bayat

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  1. Somayeh Ebrahimi-Barough
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  2. Elham Hoveizi
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Correspondence to Somayeh Ebrahimi-Barough or Jafar Ai.

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Ebrahimi-Barough, S., Hoveizi, E., Yazdankhah, M. et al. Inhibitor of PI3K/Akt Signaling Pathway Small Molecule Promotes Motor Neuron Differentiation of Human Endometrial Stem Cells Cultured on Electrospun Biocomposite Polycaprolactone/Collagen Scaffolds. Mol Neurobiol 54, 2547–2554 (2017). https://doi.org/10.1007/s12035-016-9828-z

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