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Cellular and Molecular Neurobiology

, Volume 37, Issue 2, pp 275–289 | Cite as

Motor Neuron Transdifferentiation of Neural Stem Cell from Adipose-Derived Stem Cell Characterized by Differential Gene Expression

  • Marzieh Darvishi
  • Taki Tiraihi
  • Seyed A. Mesbah-Namin
  • AliReza Delshad
  • Taher Taheri
Original Research

Abstract

Adipose-derived stem cells (ADSC) are adult stem cells which can be induced into motor neuron-like cells (MNLC) with a preinduction-induction protocol. The purpose of this study is to generate MNLC from neural stem cells (NSC) derived from ADSC. The latter were isolated from the perinephric regions of Sprague–Dawley rats, transdifferentiated into neurospheres (NS) using B27, EGF, and bFGF. After generating NSC from the NS, they induced into MNLC by treating them with Shh and RA, then with GDNF, CNTF, BDNF, and NT-3. The ADSC lineage was evaluated by its mesodermal differentiation and was characterized by immunostaining with CD90, CD105, CD49d, CD106, CD31, CD45, and stemness genes (Oct4, Nanog, and Sox2). The NS and the NSC were evaluated by immunostaining with nestin, NF68, and Neurod1, while the MNLC were evaluated by ISLET1, Olig2, and HB9 genes. The efficiency of MNLC generation was more than 95 ± 1.4 % (mean ± SEM). The in vitro generated myotubes were innervated by the MNLC. The induced ADSC adopted multipolar motor neuron morphology, and they expressed ISLET1, Olig2, and HB9. We conclude that ADSC can be induced into motor neuron phenotype with high efficiency, associated with differential expression of the motor neuron gene. The release of MNLC synaptic vesicles was demonstrated by FM1-43, and they were immunostained with synaptophysin. This activity was correlated with the intracellular calcium ion shift and membrane depolarization upon stimulation as was demonstrated by the calcium indicator and the voltage-sensitive dye, respectively.

Keywords

Stem cell ADSC Motor neurons Transdifferentiation Induction 

Notes

Acknowledgments

The project was funded by Shefa Neurosciences Research Center at Khatam Al-Anbia Hospital, Tehran, Iran (Grant# 86-N-105). We are also grateful for the support of the Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. We would like express our deep gratitude for Mrs. HH AliAkbar for editing the manuscript.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Marzieh Darvishi
    • 1
  • Taki Tiraihi
    • 1
  • Seyed A. Mesbah-Namin
    • 2
  • AliReza Delshad
    • 3
  • Taher Taheri
    • 4
  1. 1.Department of Anatomical Sciences, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran
  2. 2.Department of Clinical Biochemistry, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran
  3. 3.Department of AnatomyShahed UniversityTehranIran
  4. 4.Shefa Neurosciences Research CenterKhatam Al-Anbia HospitalTehranIran

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