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.
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References
Abdanipour A, Tiraihi T (2012) Induction of adipose-derived stem cell into motoneuron-like cells using selegiline as preinducer. Brain Res 1440:23–33. doi:10.1016/j.brainres.2011.12.051
Aleksandrova MA, Saburina IN, Poltavtseva RA, Revishchin AV, Korochkin LI, Sukhikh GT (2002) Behavior of human neural progenitor cells transplanted to rat brain. Brain Res Dev Brain Res 134(1–2):143–148
Apati A, Paszty K, Hegedus L, Kolacsek O, Orban TI, Erdei Z, Szebenyi K, Pentek A, Enyedi A, Sarkadi B (2013) Characterization of calcium signals in human embryonic stem cells and in their differentiated offspring by a stably integrated calcium indicator protein. Cell Signal 25(4):752–759. doi:10.1016/j.cellsig.2012.12.024
Asuelime GE, Shi Y (2012) A case of cellular alchemy: lineage reprogramming and its potential in regenerative medicine. J Mol Cell Biol 4(4):190–196. doi:10.1093/jmcb/mjs005
Bamford NS, Robinson S, Palmiter RD, Joyce JA, Moore C, Meshul CK (2004) Dopamine modulates release from corticostriatal terminals. J Neurosci 24(43):9541–9552. doi:10.1523/jneurosci.2891-04.2004
Baptista LS, Silva KR, Pedrosa CS, Amaral RJ, Belizario JV, Borojevic R, Granjeiro JM (2013) Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells: a comparison with human adipose-derived mesenchymal stromal cells. Artif Organs 37(12):1068–1075. doi:10.1111/aor.12121
Bieberich E, Anthony GE (2004) Neuronal differentiation and synapse formation of PC12 and embryonic stem cells on interdigitated microelectrode arrays: contact structures for neuron-to-electrode signal transmission (NEST). Biosens Bioelectron 19(8):923–931
Canepari M, Zecevic D (2010) Membrane potential imaging in the nervous system: methods and applications. Springer, New York
Cardozo A, Ielpi M, Gomez D, Argibay P (2010) Differential expression of Shh and BMP signaling in the potential conversion of human adipose tissue stem cells into neuron-like cells in vitro. Gene Expr 14(6):307–319
Chemla S, Chavane F (2010) Voltage-sensitive dye imaging: technique review and models. J Physiol Paris 104(1–2):40–50. doi:10.1016/j.jphysparis.2009.11.009
Darabi S, Tiraihi T, Ruintan A, Abbaszadeh HA, Delshad A, Taheri T (2013) Polarized neural stem cells derived from adult bone marrow stromal cells develop a rosette-like structure. Vitro Cell Dev Biol Anim 49(8):638–652. doi:10.1007/s11626-013-9628-y
Dawitz J, Kroon T, Hjorth JJ, Meredith RM (2011) Functional calcium imaging in developing cortical networks. J Vis Exp. doi:10.3791/3550
de Peppo GM, Marolt D (2012) State of the art in stem cell research: human embryonic stem cells, induced pluripotent stem cells, and transdifferentiation. J Blood Transfus 2012:317632. doi:10.1155/2012/317632
Dessaud E, Yang LL, Hill K, Cox B, Ulloa F, Ribeiro A, Mynett A, Novitch BG, Briscoe J (2007) Interpretation of the sonic hedgehog morphogen gradient by a temporal adaptation mechanism. Nature 450(7170):717–720. doi:10.1038/nature06347
Gharibani PM, Tiraihi T, Arabkheradmand J (2010) In vitro differentiation of GABAergic cells from bone marrow stromal cells using potassium chloride as inducer. Restor Neurol Neurosci 28(3):367–377. doi:10.3233/rnn-2010-0539
Ghorbanian MT, Tiraihi T, Mesbah-Namin SA, Fathollahi Y (2010) Selegiline is an efficient and potent inducer for bone marrow stromal cell differentiation into neuronal phenotype. Neurol Res 32(2):185–193. doi:10.1179/174313209x409016
Gonzalez-Garza MT, Martinez HR, Caro-Osorio E, Cruz-Vega DE, Hernandez-Torre M, Moreno-Cuevas JE (2013) Differentiation of CD133+stem cells from amyotrophic lateral sclerosis patients into preneuron cells. Stem Cells Transl Med 2(2):129–135. doi:10.5966/sctm.2012-0077
Graf T (2011) Historical origins of transdifferentiation and reprogramming. Cell Stem Cell 9(6):504–516. doi:10.1016/j.stem.2011.11.012
Griesinger CB, Richards CD, Ashmore JF (2002) Fm1-43 reveals membrane recycling in adult inner hair cells of the mammalian cochlea. J Neurosci 22(10):3939–3952.
Guo X, Johe K, Molnar P, Davis H, Hickman J (2010) Characterization of a human fetal spinal cord stem cell line, NSI-566RSC, and its induction to functional motoneurons. J Tissue Eng Regen Med 4(3):181–193. doi:10.1002/term.223
Henkel AW, Lubke J, Betz WJ (1996) FM1-43 dye ultrastructural localization in and release from frog motor nerve terminals. Proc Natl Acad Sci USA 93(5):1918–1923
Hester ME, Murtha MJ, Song S, Rao M, Miranda CJ, Meyer K, Tian J, Boulting G, Schaffer DV, Zhu MX (2011) Rapid and efficient generation of functional motor neurons from human pluripotent stem cells using gene delivered transcription factor codes. Mol Ther 19(10):1905–1912
Hu BY, Zhang SC (2010) Directed differentiation of neural-stem cells and subtype-specific neurons from hESCs. Methods Mol Biol 636:123–137. doi:10.1007/978-1-60761-691-7_8
Hu W, Guan FX, Li Y, Tang YJ, Yang F, Yang B (2013) New methods for inducing the differentiation of amniotic-derived mesenchymal stem cells into motor neuron precursor cells. Tissue Cell 45(5):295–305. doi:10.1016/j.tice.2013.03.002
Joo KM, Jin J, Kang BG, Lee SJ, Kim KH, Yang H, Lee Y-A, Cho YJ, Im Y-S, Lee D-S (2012) Trans-differentiation of neural stem cells: a therapeutic mechanism against the radiation induced brain damage. PLoS ONE 7(2):e25936
Kokai LE, Rubin JP, Marra KG (2005) The potential of adipose-derived adult stem cells as a source of neuronal progenitor cells. Plast Reconstr Surg 116(5):1453–1460
Krabbe C, Zimmer J, Meyer M (2005) Neural transdifferentiation of mesenchymal stem cells: a critical review. APMIS 113(11–12):831–844. doi:10.1111/j.1600-0463.2005.apm_3061.x
Leao RN, Reis A, Emirandetti A, Lewicka M, Hermanson O, Fisahn A (2010) A voltage-sensitive dye-based assay for the identification of differentiated neurons derived from embryonic neural stem cell cultures. PLoS ONE 5(11):e13833. doi:10.1371/journal.pone.0013833
Lee HJ, Kim KS, Ahn J, Bae HM, Lim I, Kim SU (2014) Human motor neurons generated from neural stem cells delay clinical onset and prolong life in ALS mouse model. PLoS ONE 9(5):e97518. doi:10.1371/journal.pone.0097518
Lepore AC, Maragakis NJ (2007) Targeted stem cell transplantation strategies in ALS. Neurochem Int 50(7–8):966–975. doi:10.1016/j.neuint.2006.09.005
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):370–373. doi:10.1097/WNR.0b013e3283469615
Miles GB, Yohn DC, Wichterle H, Jessell TM, Rafuse VF, Brownstone RM (2004) Functional properties of motoneurons derived from mouse embryonic stem cells. J Neurosci 24(36):7848–7858. doi:10.1523/jneurosci.1972-04.2004
Mirakhori F, Zeynali B, Salekdeh GH, Baharvand H (2014) Induced neural lineage cells as repair kits: so close, yet so far away. J Cell Physiol 229(6):728–742. doi:10.1002/jcp.24509
Mohammad-Gharibani P, Tiraihi T, Mesbah-Namin SA, Arabkheradmand J, Kazemi H (2012) Induction of bone marrow stromal cells into GABAergic neuronal phenotype using creatine as inducer. Restor Neurol Neurosci 30(6):511–525. doi:10.3233/rnn-2012-100155
Monni E, Congiu T, Massa D, Nat R, Diana A (2011) Human neurospheres: from stained sections to three-dimensional assembly. Transl Neurosci 2(1):43–48
Naghdi M, Tiraihi T, Namin SA, Arabkheradmand J (2009) Transdifferentiation of bone marrow stromal cells into cholinergic neuronal phenotype: a potential source for cell therapy in spinal cord injury. Cytotherapy 11(2):137–152. doi:10.1080/14653240802716582
Nakanishi S, Okazawa M (2006) Membrane potential-regulated Ca2 + signalling in development and maturation of mammalian cerebellar granule cells. J Physiol 575(Pt 2):389–395. doi:10.1113/jphysiol.2006.113340
Nizzardo M, Simone C, Falcone M, Locatelli F, Riboldi G, Comi GP, Corti S (2010) Human motor neuron generation from embryonic stem cells and induced pluripotent stem cells. Cell Mol Life Sci 67(22):3837–3847. doi:10.1007/s00018-010-0463-y
Petros TJ, Tyson JA, Anderson SA (2011) Pluripotent stem cells for the study of CNS development. Front Mol Neurosci 4:30. doi:10.3389/fnmol.2011.00030
Rui Y, Xu L, Chen R, Zhang T, Lin S, Hou Y, Liu Y, Meng F, Liu Z, Ni M, Tsang KS, Yang F, Wang C, Chan HC, Jiang X, Li G (2015) Epigenetic memory gained by priming with osteogenic induction medium improves osteogenesis and other properties of mesenchymal stem cells. Sci Rep 5:11056. doi:10.1038/srep11056
Silani V, Fogh I, Ratti A, Sassone J, Ciammola A, Cova L (2002) Stem cells in the treatment of amyotrophic lateral sclerosis (ALS). Amyotroph Lateral Scler Other Motor Neuron Disord 3(4):173–181
Son EY, Ichida JK, Wainger BJ, Toma JS, Rafuse VF, Woolf CJ, Eggan K (2011) Conversion of mouse and human fibroblasts into functional spinal motor neurons. Cell Stem Cell 9(3):205–218. doi:10.1016/j.stem.2011.07.014
Staple JK, Osen-Sand A, Benfenati F, Pich EM, Catsicas S (1997) Molecular and functional diversity at synapses of individual neurons in vitro. Eur J Neurosci 9(4):721–731
Su H, Zhang W, Yang X, Qin D, Sang Y, Wu C, Wong WM, Yuan Q, So KF, Wu W (2012) Neural progenitor cells generate motoneuron-like cells to form functional connections with target muscles after transplantation into the musculocutaneous nerve. Cell Transplant 21(12):2651–2663. doi:10.3727/096368912x654975
Sudhof TC (2012) Calcium control of neurotransmitter release. Cold Spring Harb Perspect Biol 4(1):a011353. doi:10.1101/cshperspect.a011353
Wichterle H, Peljto M (2008) Differentiation of mouse embryonic stem cells to spinal motor neurons. Curr Protoc Stem Cell Biol Chapter 1 Unit 1H 1 1-1H 1 9. doi:10.1002/9780470151808.sc01h01s5
Wu CY, Whye D, Mason RW, Wang W (2012) Efficient differentiation of mouse embryonic stem cells into motor neurons. J Vis Exp 64:e3813. doi:10.3791/3813
Yaghoobi MM, Mowla SJ (2006) Differential gene expression pattern of neurotrophins and their receptors during neuronal differentiation of rat bone marrow stromal cells. Neurosci Lett 397(1–2):149–154. doi:10.1016/j.neulet.2005.12.009
Yoo J, Kim HS, Hwang DY (2013) Stem cells as promising therapeutic options for neurological disorders. J Cell Biochem 114(4):743–753. doi:10.1002/jcb.24427
Zhou Q, Melton DA (2008) Extreme makeover: converting one cell into another. Cell Stem Cell 3(4):382–388. doi:10.1016/j.stem.2008.09.015
Zurn AD, Winkel L, Menoud A, Djabali K, Aebischer P (1996) Combined effects of GDNF, BDNF, and CNTF on motoneuron differentiation in vitro. J Neurosci Res 44(2):133–141. doi:10.1002/(SICI)1097-4547(19960415)44:2<133:AID-JNR5>3.0.CO;2-E
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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Darvishi, M., Tiraihi, T., Mesbah-Namin, S.A. et al. Motor Neuron Transdifferentiation of Neural Stem Cell from Adipose-Derived Stem Cell Characterized by Differential Gene Expression. Cell Mol Neurobiol 37, 275–289 (2017). https://doi.org/10.1007/s10571-016-0368-x
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DOI: https://doi.org/10.1007/s10571-016-0368-x