Molecular Neurobiology

, Volume 52, Issue 3, pp 1704–1713 | Cite as

Evaluation of Motor Neuron-Like Cell Differentiation of hEnSCs on Biodegradable PLGA Nanofiber Scaffolds

  • Somayeh Ebrahimi-Barough
  • Abbas Norouzi Javidan
  • Hoshangh Saberi
  • Mohammad Tghi Joghataei
  • Reza Rahbarghazi
  • Esmaeil Mirzaei
  • Faezeh Faghihi
  • Sadegh Shirian
  • Armin Ai
  • Jafar Ai


Human endometrium is a high-dynamic tissue that contains human endometrial stem cells (hEnSCs) which can be differentiated into a number of cell lineages. The differentiation of hEnSCs into many cell lineages such as osteoblast, adipocyte, and neural cells has been investigated previously. However, the differentiation of these stem cells into motor neuron-like cells has not been investigated yet. Different biochemical and topographical cues can affect the differentiation of stem cells into a specific cell. The aim of this study was to investigate the capability of hEnSCs to be differentiated into motor neuron-like cells under biochemical and topographical cues. The biocompatible and biodegradable poly(lactic-co-glycolic acid) (PLGA) electrospun nanofibrous scaffold was used as a topographical cue. Human EnSCs were cultured on the PLGA scaffold and tissue culture polystyrene (TCP), then differentiation of hEnSCs into motor neuron-like cells under induction media including retinoic acid (RA) and sonic hedgehog (Shh) were evaluated for 15 days. The proliferation rate of cells was assayed by using MTT assay. The morphology of cells was studied by scanning electron microscopy imaging, and the expression of motor neuron-specific markers by real-time PCR and immunocytochemistry. Results showed that survival and differentiation of hEnSCs into motor neuron-like cells on the PLGA scaffold were better than those on the TCP group. Taken together, the results suggest that differentiated hEnSCs on PLGA can provide a suitable, three-dimensional situation for neuronal survival and outgrowth for regeneration of the central nervous system, and these cells may be a potential candidate in cellular therapy for motor neuron diseases.


Human endometrial stem cell Motor neuron differentiation Nanofibrous scaffolds PLGA 



We thank the Iran National Science Foundation (INSF) for the financial support (grant number 92004774) and Tehran University of Medical Sciences for this research.


  1. 1.
    Parr AM, Kulbatski I, Zahir T, Wang X, Yue C, Keating A, Tator CH (2008) Transplanted adult spinal cord-derived neural stem/progenitor cells promote early functional recovery after rat spinal cord injury. Neuroscience 155(3):760–770CrossRefPubMedGoogle Scholar
  2. 2.
    Yuan YM, He C (2013) The glial scar in spinal cord injury and repair. Neurosci Bull 29(4):421–435CrossRefPubMedGoogle Scholar
  3. 3.
    Rolls A, Shechter R, Schwartz M (2009) The bright side of the glial scar in CNS repair. Nat Rev Neurosci 10(3):235–241CrossRefPubMedGoogle Scholar
  4. 4.
    Ronsyn MW, Berneman ZN, Van Tendeloo VFI, Jorens PG, Ponsaerts P (2008) Can cell therapy heal a spinal cord injury? Spinal Cord 46(8):532–539CrossRefPubMedGoogle Scholar
  5. 5.
    Kim BG, Hwang DH, Lee SI, Kim EJ, Kim SU (2007) Stem cell-based cell therapy for spinal cord injury. Cell Transplant 16(4):355–364CrossRefPubMedGoogle Scholar
  6. 6.
    Mothe AJ, Tator CH (2013) Review of transplantation of neural stem/progenitor cells for spinal cord injury. Int J Dev Neurosci 31(7):701–713CrossRefPubMedGoogle Scholar
  7. 7.
    Watanabe K, Nakamura M, Iwanami A, Fujita Y, Kanemura Y, Toyama Y, Okano H (2004) Comparison between fetal spinal-cord- and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury. Dev Neurosci 26(2–4):275–287PubMedGoogle Scholar
  8. 8.
    Yazdani SO, Pedram M, Hafizi M, Kabiri M, Soleimani M, Dehghan MM, Jahanzad I, Gheisari Y, Hashemi SM (2012) A comparison between neurally induced bone marrow derived mesenchymal stem cells and olfactory ensheathing glial cells to repair spinal cord injuries in rat. Tissue Cell 44(4):205–213CrossRefPubMedGoogle Scholar
  9. 9.
    Kang CE, Poon PC, Tator CH, Shoichet MS (2009) A new paradigm for local and sustained release of therapeutic molecules to the injured spinal cord for neuroprotection and tissue repair. Tissue Eng Part A 15(3):595–604CrossRefPubMedGoogle Scholar
  10. 10.
    Piantino J, Burdick JA, Goldberg D, Langer R, Benowitz LI (2006) An injectable, biodegradable hydrogel for trophic factor delivery enhances axonal rewiring and improves performance after spinal cord injury. Exp Neurol 201(2):359–367CrossRefPubMedGoogle Scholar
  11. 11.
    Ghoroghi MF, Beygom Hejazian L, Esmaielzade B, Dodel M, Roudbari M, Nobakht M (2013) Evaluation of the effect of NT-3 and biodegradable poly-L-lactic acid nanofiber scaffolds on differentiation of rat hair follicle stem cells into neural cells in vitro. J Mol Neurosci 51(3):318–327CrossRefGoogle Scholar
  12. 12.
    Li WJ, Laurencin CT, Caterson EJ, Tuan RS, Ko FK (2002) Electrospun nanofibrous structure: a novel scaffold for tissue engineering. J Biomed Mater Res 60(4):613–621CrossRefPubMedGoogle Scholar
  13. 13.
    Lim SH, Mao HQ (2009) Electrospun scaffolds for stem cell engineering. Adv Drug Deliv Rev 61(12):1084–1096CrossRefPubMedGoogle Scholar
  14. 14.
    Ai J, Kiasat-Dolatabadi A, Ebrahimi-Barough S, Ai A, Lotfibakhshaiesh N, Norouzi-Javidan A, Saberi H, Arjmand B, Aghayan HR (2013) Polymeric scaffolds in neural tissue engineering: a review. Arch Neurosci 1(1):15–20CrossRefGoogle Scholar
  15. 15.
    Hsu SH, Chang CJ, Tang CM, Lin FT (2004) In vitro and in vivo effects of Ginkgo biloba extract EGb 761 on seeded Schwann cells within poly(DL-lactic acid-co-glycolic acid) conduits for peripheral nerve regeneration. J Biomater Appl 19(2):163–182CrossRefPubMedGoogle Scholar
  16. 16.
    Hou SY, Zhang Y, Quan DP, Liu XL, Zhu JK (2006) Tissue-engineered peripheral nerve grafting by differentiated bone marrow stromal cells. Neuroscience 140(1):101–110CrossRefPubMedGoogle Scholar
  17. 17.
    Wen X, Tresco PA (2006) Fabrication and characterization of permeable degradable poly(DL-lactide-co-glycolide) (PLGA) hollow fiber phase inversion membranes for use as nerve tract guidance channels. Biomaterials 27(20):3800–3809CrossRefPubMedGoogle Scholar
  18. 18.
    Ulrich D, Muralitharan R, Gargett CE (2013) Toward the use of endometrial and menstrual blood mesenchymal stem cells for cell-based therapies. Expert Opin Biol Ther 13(10):1387–1400CrossRefPubMedGoogle Scholar
  19. 19.
    Gargett CE, Schwab KE, Zillwood RM, Nguyen HP, Wu D (2009) Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium. Biol Reprod 80(6):1136–1145CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Wolff EF, Gao XB, Yao KV, Andrews ZB, Du H, Elsworth JD, Taylor HS (2011) Endometrial stem cell transplantation restores dopamine production in a Parkinson's disease model. J Cell Mol Med 15(4):747–755CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Tavakol S, Aligholi H, Gorji A, Eshaghabadi A, Hoveizi E, Tavakol B, Rezayat SM, Ai J (2014) Thermogel nanofiber induces human endometrial-derived stromal cells to neural differentiation: in vitro and in vivo studies in rat. J Biomed Mater Res A. doi: 10.1002/jbm.a.35117 Google Scholar
  22. 22.
    Navaei-Nigjeh M, Amoabedini G, Noroozi A, Azami M, Asmani MN, Ebrahimi-Barough S, Saberi H, Ai A, Ai J (2014) Enhancing neuronal growth from human endometrial stem cells derived neuron-like cells in three-dimensional fibrin gel for nerve tissue engineering. J Biomed Mater Res A 102(8):2533–2543CrossRefPubMedGoogle Scholar
  23. 23.
    Mobarakeh ZT, Ai J, Yazdani F, Sorkhabadi SM, Ghanbari Z, Javidan AN, Mortazavi- Tabatabaei SA, Massumi M, Barough SE (2012) Human endometrial stem cells as a new source for programming to neural cells. Cell Boil Inter Rep 19(1):7–14CrossRefGoogle Scholar
  24. 24.
    Ebrahimi-Barough S, Kouchesfahani HM, Ai J, Massumi M (2013) Differentiation of human endometrial stromal cells into oligodendrocyte progenitor cells (OPCs). J Mol Neurosci 51(2):265–273CrossRefPubMedGoogle Scholar
  25. 25.
    Ebrahimi-Barough S, Kouchesfehani HM, Ai J, Mahmoodinia M, Tavakol S, Massumi M (2013) Programming of human endometrial-derived stromal cells (EnSCs) into pre-oligodendrocyte cells by overexpression of miR-219. Neurosci Lett 537:65–70CrossRefPubMedGoogle Scholar
  26. 26.
    Ebrahimi-Barough S, Massumi M, Kouchesfahani HM, Ai J (2013) Derivation of pre-oligodendrocytes from human endometrial stromal cells by using overexpression of microRNA 338. J Mol Neurosci 51(2):337–343CrossRefPubMedGoogle Scholar
  27. 27.
    Asmani MN, Ai J, Amoabediny G, Noroozi A, Azami M, Ebrahimi-Barough S, Navaei-Nigjeh M, Ai A, Jafarabadi M (2013) Three-dimensional culture of differentiated endometrial stromal cells to oligodendrocyte progenitor cells (OPCs) in fibrin hydrogel. Cell Boil Int 37(12):1340–1349CrossRefGoogle Scholar
  28. 28.
    Ai J, Shahverdi AR, Barough SE, Kouchesfehani HM, Heidari S, Roozafzoon R, Verdi J, Khoshzaban A (2012) Derivation of adipocytes from human endometrial stem cells (EnSCs). J Reprod Infertile 13(3):151–157Google Scholar
  29. 29.
    Azami M, Ai J, Ebrahimi-Barough S, Farokhi M, Fard SE (2013) In vitro evaluation of biomimetic nanocomposite scaffold using endometrial stem cell derived osteoblast-like cells. Tissue Cell 45(5):328–337CrossRefPubMedGoogle Scholar
  30. 30.
    Ai J, Azizi E, Shamsian A, Eslami A, Khoshzaban A, Ebrahimi-Barough S, Ai A, Alizadeh A (2014) BMP-2 can promote the osteogenic differentiation of human endometrial stem cells. ABM 8:21–29Google Scholar
  31. 31.
    Niknamasl A, Ostad SN, Soleimani M, Azami M, Salmani MK, Lotfibakhshaiesh N, Ebrahimi-Barough S, Karimi R, Roozafzoon R, Ai J (2014) A new approach for pancreatic tissue engineering: human endometrial stem cells encapsulated in fibrin gel can differentiate to pancreatic islet beta-cell. Cell Biol Int 6: doi:  10.1002/cbin.10314
  32. 32.
    Liqing Y, Jia G, Jiqing C, Ran G, Fei C, Jie K, Yanyun W, Cheng Z (2011) Directed differentiation of motor neuron cell-like cells from human adipose-derived stem cells in vitro. Neuroreport 22(8):370–373CrossRefPubMedGoogle Scholar
  33. 33.
    Chan RW, Schwab KE, Gargett CE (2004) Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod 70(6):1738–1750CrossRefPubMedGoogle Scholar
  34. 34.
    Gargett CE, Nguyen HP, Ye L (2012) Endometrial regeneration and endometrial stem/progenitor cells. Rev Endocr Metab Disord 13(4):235–251CrossRefPubMedGoogle Scholar
  35. 35.
    Wang H, Jin P, Sabatino M, Ren J, Civini S, Bogin V, Ichim TE, Stroncek DF (2012) Comparison of endometrial regenerative cells and bone marrow stromal cells. J Transl Med 10(1):207–213CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Ouyang Y, Huang C, Zhu Y, Fan C, Ke Q (2013) Fabrication of seamless electrospun collagen/PLGA conduits whose walls comprise highly longitudinal aligned nanofibers for nerve regeneration. J Biomed Nanotechnol 9(6):931–943CrossRefPubMedGoogle Scholar
  37. 37.
    Huimin Y, Lei Z, Yunfang W, Feng L, Lidong G, Shaoqing L, Fang Y, Xue N, Cixian B, Feng L, Yongnian Y, Xuetao P (2006) Proliferation and differentiation into endothelial cells of human bone marrow mesenchymal stem cells (MSCs) on poly DL-lactic-co-glycolic acid (PLGA) films. Sci Bull 51(1):1328–1333Google Scholar
  38. 38.
    Park HW, Cho JS, Park CK, Jung SJ, Park CH, Lee SJ, Oh S, Park YS, Chang MS (2012) Directed induction of functional motor neuron-like cells from genetically engineered human mesenchymal stem cells. PLoS ONE 7(4):35–44CrossRefGoogle Scholar
  39. 39.
    Li XJ, Hu BY, Jones SA, Zhang YS, Lavaute T, Du ZW, Zhang SC (2008) Directed differentiation of ventral spinal progenitors and motor neurons from human embryonic stem cells by small molecules. Stem Cells 26(4):886–893CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Ericson J, Morton S, Kawakami A, Roelink H, Jessell TM (1996) Two critical periods of sonic hedgehog signaling required for the specification of motor neuron identity. Cell 87(4):661–673CrossRefPubMedGoogle Scholar
  41. 41.
    Ericson J, Rashbass P, Sched A, Brenner-Morton S, Kawakami A, Heyningen V, Jessell TM, Briscoe J (1997) Pax6 controls progenitor cell identity and neuronal fate in response to graded Shh signaling. Cell 90(4):169–180CrossRefPubMedGoogle Scholar
  42. 42.
    Hendrickson ML, Rao AJ, Demerdash ON, Kalil RE (2011) Expression of nestin by neural cells in the adult rat and human brain. PLoS ONE 6:1–15CrossRefGoogle Scholar
  43. 43.
    Gillette BM, Rossen NS, Das N, Leong D, Wang M, Dugar A, Sia SK (2011) Engineering extracellular matrix structure in 3D multiphase tissues. Biomaterials 32(32):8067–8076CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Li J, Tao R, Wu W, Cao H, Xin J, Li J, Guo J, Jiang L, Gao C, Demetriou AA, Farkas DL, Li L (2010) 3D PLGA scaffolds improve differentiation and function of bone marrow mesenchymal stem cell-derived hepatocytes. Stem Cells Dev 19(32):1427–1436CrossRefPubMedGoogle Scholar
  45. 45.
    Wang J, Ye R, Wei Y, Wang H, Xu X, Zhang F, Qu J, Zuo B, Zhang H (2012) The effects of electrospun TSF nanofiber diameter and alignment on neuronal differentiation of human embryonic stem cells. J Biomed Mater Res A 100(9):632–645CrossRefPubMedGoogle Scholar
  46. 46.
    Binan L, Tendey C, De G, Crescenzo R, Ayoubi E, Ajji A, Jolicoeur M (2014) Differentiation of neuronal stem cells into motor neurons using electrospun poly-L-lactic acid/gelatin scaffold. Biomaterials 35(2):664–6674CrossRefPubMedGoogle Scholar
  47. 47.
    Prabhakaran MP, Venugopal JR, Chyan TT, Hai LB, Chan CK, Lim AY, Ramakrishna S (2008) Electrospun biocomposite nanofibrous scaffolds for neural tissue engineering. Tissue Eng Part A 14(11):1787–1797CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Somayeh Ebrahimi-Barough
    • 1
    • 2
  • Abbas Norouzi Javidan
    • 1
  • Hoshangh Saberi
    • 1
  • Mohammad Tghi Joghataei
    • 3
    • 4
  • Reza Rahbarghazi
    • 5
  • Esmaeil Mirzaei
    • 6
  • Faezeh Faghihi
    • 3
  • Sadegh Shirian
    • 1
  • Armin Ai
    • 7
  • Jafar Ai
    • 1
    • 2
  1. 1.Brain and Spinal Cord Injury Research Center (BASIR)Tehran University of Medical SciencesTehranIran
  2. 2.Department of Tissue Engineering, Faculty of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran
  3. 3.Cellular and Molecular Research CenterIran University of Medical SciencesTehranIran
  4. 4.Department of Anatomy, School of MedicineIran University of Medical SciencesTehranIran
  5. 5.Umbilical Cord Stem Cell Research CenterTabriz University of Medical SciencesTabrizIran
  6. 6.Department of Medical Nanotechnology, School of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran
  7. 7.Dentistry FacultyTehran University of Medical SciencesTehranIran

Personalised recommendations