Biomedicine and Biotechnology
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Biomedicine and Biotechnology. 2014, 2(4), 66-79
DOI: 10.12691/bb-2-4-2
Open AccessArticle

Transdifferentiation of Bone Marrow Mesenchymal Stem Cells into Neural Cells via Cerebrospinal Fluid

Dina Y. Otify1, 2, , EmanA. Youssef1, Naglaa B. Nagy2, Mona K. Marei2 and Magda I. Youssif1

1Department of Histochemistry and Cell Biology, Medical Research Institute, Alexandria University, Egypt

2Tissue Engineering Laboratories, Faculty of Dentistry, Alexandria University, Egypt

Pub. Date: November 21, 2014

Cite this paper:
Dina Y. Otify, EmanA. Youssef, Naglaa B. Nagy, Mona K. Marei and Magda I. Youssif. Transdifferentiation of Bone Marrow Mesenchymal Stem Cells into Neural Cells via Cerebrospinal Fluid. Biomedicine and Biotechnology. 2014; 2(4):66-79. doi: 10.12691/bb-2-4-2

Abstract

Research showed that adult bone marrow mesenchymal stem cells (BM-MSCs) differentiate into mesodermal cell types and also reprogram to transdifferentiate into endodermal and ectodermal cell types. The present study induced BM-MSCs to transdifferentiate into neural-like cells (either neurons or glial cells) using Cerebrospinal fluid(CSF)in vitro. The harvested BM-MSCs from rabbit femur were cultured and characterized immunocytochemically by CD146 that showed positive reaction for MSCs. Autologous CSF was added daily to the culture media. Dramatic morphological and cytochemical modifications were observed in the cultured cells and identified by different staining methods. Nissl bodies observed in the soma usingcresyl violet stain, and cell bodies' processes (axons and dendrites) usingsilver impregnation, proved that BM-MSCs differentiated into neuronal cells. Periodic Acid-Schiff demonstrated glycogen granules in astrocytes used to verify astrocytes differentiation. Immunocytochemical examination for glial fibrillic acid protein marker showed positive reaction indicating successful astrocytes differentiation. This study showed that induction of BM-MSCs with CSF mimics the strategy done in CNS. CSFcould provide an essential niche for promoting the transdifferentiation of BM-MSCs into neural cells, that hopefully help in treating acute and chronic neurodegenerative diseases.

Keywords:
BM-MSCs CSF neuronal cells astrocytes cresyl violet silver impregnation Periodic Acid-Schiff CD146 glial fibrillic acid protein

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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References:

[1]  Bredesen DE, Rao RV, Mehlen P. Cell death in the nervous system.Nature 2006 19; 443(7113): 796-802.
 
[2]  Chen MB, Zhang F, Lineaweaver WC. Luminal fillers in nerve conduits for peripheral nerve repair. Ann PlastSurg 2006b; 57(4): 462-71.
 
[3]  Martino S, D'Angelo F, Armentano I, Kenny JM, Orlacchio A. Stem cell-biomaterial interactions for regenerative medicine. BiotechnolAdv 2012; 30(1): 338-51.
 
[4]  Zhang W, Zhang F, Shi H, Tan R, Han S, Ye G, Pan S, Sun F, Liu X. Comparisons of rabbit bone marrow mesenchymal stem cell isolation and culture methods invitro. PLoS ONE 2014; 9(2): e88794.
 
[5]  Krabbe C, Zimmer J, Meyer M. Neural transdifferentiation of mesenchymal stem cells- a critical review. APMIS 2005; 113(11-12): 831-44.
 
[6]  Jungebluth P, Alici E, Baiguera S, Le Blanc K, Blomberg P, Bozóky B, Crowley C, Einarsson O, Grinnemo KH, Gudbjartsson T, Le Guyader S, Henriksson G, Hermanson O, Juto JE, Leidner B, Lilja T, Liska J, Luedde T, Lundin V, Moll G, Nilsson B, Roderburg C, Strömblad S, Sutlu T, Teixeira AI, Watz E, Seifalian A, Macchiarini P. Tracheobronchial transplantation with a stem-cell-seeded bioartificialnanocomposite: a proof-of-concept study. Lancet 2011; 378(9808):1997-2004.
 
[7]  Heino TJ, Hentunen TA. Differentiation of osteoblasts and osteocytes from mesenchymal stem cells.Curr Stem Cell Res Ther 2008; 3(2): 131–145.
 
[8]  Karagianni M, Brinkmann I, Kinzebach S, Grassl M, Weiss C, Bugert P, Bieback. A comparative analysis of the adipogenic potential in human mesenchymal stromal cells from cord blood and other sources.Cytotherapy 2013; 15(1):76-88.
 
[9]  Catacchio I, Berardi S, Reale A, De Luisi A, Racanelli V, Vacca A, Ria R. Evidence for Bone Marrow Adult Stem Cell Plasticity: Properties, Molecular Mechanisms, Negative Aspects, and Clinical Applications of Hematopoietic and Mesenchymal Stem Cells Transdifferentiation. Stem Cells Int 2013; 2013: 589139.
 
[10]  Hermann A, Liebau S, Gastl R, Fickert S, Habisch HJ, Fiedler J, Schwarz J, Brenner R, Storch A Comparative analysis of neuroectodermal differentiation capacity of human bone marrow stromal cells using various conversion protocols. J Neurosci Res. 2006; 83(8): 1502-14.
 
[11]  Zeng R, Wang LW, Hu ZB, Gou WT, Wei JS, Lin H, Sun X, Chen LX, Yang LJ. Differentiation of human bone marrow mesenchymal stem cells into neuron-like cells in vitro. Spine (Phila Pa 1976) 2011; 36(13): 997-1005.
 
[12]  Woodbury D, Schwarz EJ, Prockop DJ, Black IB. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 2000; 61(4):364-70.
 
[13]  Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM.Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002; 418(6893): 41-9.
 
[14]  Hermann A, Gastl R, Liebau S, Popa MO, Fiedler J, Boehm BO, Maisel M, Lerche H, Schwarz J, Brenner R, Storch A.Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells. J Cell Sci 2004; 117(Pt 19): 4411-22.
 
[15]  Qian DX, Zhang HT, Ma X, Jiang XD, Xu RX.Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells.Neurochem Res. 2010; 35(4):572-9.
 
[16]  Jiang Y, Henderson D, Blackstad M, Chen A, Miller RF, Verfaillie CM. Neuroectodermal differentiation from mouse multipotent adult progenitor cells. ProcNatlAcadSci USA 2003; 100 Suppl 1: 11854-60.
 
[17]  Wislet-Gendebien S, Hans G, Leprince P, Rigo JM, Moonen G, Rogister B. Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. Stem Cells 2005; 23(3): 392-402.
 
[18]  Sanchez-Ramos J1, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman TB, Saporta S, Janssen W, Patel N, Cooper DR, Sanberg PR.Adult bone marrow stromal cells differentiate into neural cells in vitro. ExpNeurol 2000; 164(2):247–56.
 
[19]  Mazzini L, Ferrero I, Luparello V, Rustichelli D, Gunetti M, Mareschi K, Testa L, Stecco A, Tarletti R, Miglioretti M, Fava E, Nasuelli N, Cisari C, Massara M, Vercelli R, Oggioni GD, Carriero A, Cantello R, Monaco F, Fagioli F. Mesenchymal stem cell transplantation in amyotrophic lateral sclerosis: A PHASE I clinical trial. ExpNeurol 2010; 223(1): 229-37.
 
[20]  Miyan JA, Nabiyouni M, Zendah M. Development of the brain: a vital role for cerebrospinal fluid. Can J PhysiolPharmacol 2003; 81(4): 317-28.
 
[21]  Johanson CE, Duncan JA 3rd, Klinge PM, Brinker T, Stopa EG, Silverberg GD. Multiplicity of cerebrospinal fluid functions: New challenges in health and disease. Cerebrospinal Fluid Res 2008; 5: 10.
 
[22]  Morest DK, Silver J. Precursors of neurons, neuroglia, and ependymal cells in the CNS: what are they? Where are they from? How do they get where they are going?. Glia 2003; 43(1): 6-18.
 
[23]  Ming GL, Song H. Adult neurogenesis in the mammalian brain: significant answers and significant questions. Neuron 2011; 70(4): 687-702.
 
[24]  Alcon A, CagaviBozkulak E, Qyang Y.Regenerating functional heart tissue for myocardial repair.Cell Mol Life Sci 2012; 69(16): 2635-56.
 
[25]  Eça LP, Ramalho RB, Oliveira IS, Gomes PO, Pontes P, Ferreira AT, Mazzetti MP. Comparative study of technique to obtain stem cells from bone marrow collection between the iliac crest and the femoral epiphysis in rabbits.Acta Cir Bras2009; 24(5): 400-4.
 
[26]  Marei MK, Nouh SR, Saad MM, Ismail NS. Preservation and regeneration of alveolar bone by tissue-engineered implants.Tissue Eng 2005; 11(5-6): 751-67.
 
[27]  Liu L, Duff K. A technique for serial collection of cerebrospinal fluid from the cisterna magna in mouse.J Vis Exp 2008; (21).
 
[28]  Li Y, Zhang B, Wen W, Liu S, Hao D, Liu M, Kuang HX, Huang SM.The comparison of three methods of drawing cerebrospinal fluid in rabbit.J Neurosci Methods 2012; 209(2): 398-402.
 
[29]  Ye Y, Zeng YM, Wan MR, Lu XF.Induction of human bone marrow mesenchymal stem cells, differentiation into neural-like cells using cerebrospinal fluid. Cell BiochemBiophys 2011; 59 (3): 179-84.
 
[30]  Bahadori MH, Soltani B, Heidarzadeh A, Ghasemian F, Ghadarjani G. Correlation between morphological and biological characteristics of mesenchymal stem cells and hepatocytes derived from rat mesenchymal stem cells in vitro. Afr J Biotech 2012; 11(54):11746-54.
 
[31]  Parameswaran S, Verma RS. Scanning electron microscopy preparation protocol for differentiated stem cells.Anal Biochem 2011; 416(2): 186-90.
 
[32]  Rasini V, Dominici M, Kluba T, Siegel G, Lusenti G, Northoff H,Horwitz EM, Schäfer Rl. Mesenchymal stromal/stem cells markers in the human bone marrow. Cytotherapy 2013; 15(3): 292-306.
 
[33]  Scintu F, Reali C, Pillai R, Badiali M, Sanna MA, Argiolu F, Ristaldi MS, Sogos V.Differentiation of human bone marrow stem cells into cells with a neural phenotype: diverse effects of two specific treatments. BMC Neurosci 2006; 7:14.
 
[34]  Pilati N, Barker M, Panteleimonitis S, Donga R, Hamann MA. Rapid method combining Golgi and Nissl staining to study neuronal morphology and cytoarchitecture. J HistochemCytochem 2008; 56(6):539-50.
 
[35]  Uchihara T. Silver diagnosis in neuropathology: principles, practice and revised interpretation. ActaNeuropathol 2007; 113(5):483-99.
 
[36]  Brunet JF, Allaman I, Magistretti PJ, PellerinL. Glycogen metabolism as a marker of astrocyte differentiation.J Cereb Blood Flow Metab 2010; 30(1):51-5.
 
[37]  Wislet-Gendebien S, Leprince P, Moonen G, RogisterB. Regulation of neural markers nestin and GFAP expression by cultivated bone marrow stromal cells. J Cell Sci 2003; 116(Pt 16): 3295-302.
 
[38]  Ni WF, Yin LH, Lu J, Xu HZ, Chi YL, Wu JB, et al. In vitro neural differentiation of bone marrow stromal cells induced by co-cultured olfactory ensheathing cells. NeurosciLett 2010;475(2):99-103.
 
[39]  Li N, Yang H, Lu L, Duan C, Zhao C, Zhao H. Spontaneousexpression of neural phenotype and NGF, TrkA, TrkBgenes in marrow stromal cells. BiochemBiophys ResCommun2007; 356(3): 561-568
 
[40]  Yaghoobi MM, Mahani MT. NGF and BDNF expressiondrop off in neutrally differentiated bone marrow stromal stemcells. Brain Res2008; 1203: 26-31.
 
[41]  Chen Y, Teng FY, Tang BL.Coaxing bone marrow stromal mesenchymal stem cells towards neuronal differentiation: progress and uncertainties. CellMol Life Sci. 2006; 63: 1649-57.
 
[42]  Joannides AJ, Webber DJ, Raineteau O, Kelly C, Irvine KA, Watts C, Rosser AE, Kemp PJ, Blakemore WF, Compston A, Caldwell MA, Allen ND, Chandran S. Environmental signals regulatelineage choice and temporal maturation of neural stem cells fromhuman embryonic stem cells. Brain2007; 130: 1263-75.
 
[43]  Friedenstein AJ, Chailakhyan RK, Gerasimov UV. Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet 1987; 20: 263-272.
 
[44]  Muraglia A1, Cancedda R, Quarto R. Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. J Cell Sci 2000; 113 (Pt 7): 1161-6.
 
[45]  Sacchetti B1, Funari A, Michienzi S, Di Cesare S, Piersanti S, Saggio I, Tagliafico E, Ferrari S, Robey PG, Riminucci M, Bianco P.Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 2007; 131(2): 324-336.
 
[46]  Tormin A, Li O, Brune JC, Walsh S, Schütz B, Ehinger M, Ditzel N, Kassem M, Scheding S. CD146 expression on primary nonhematopoietic bone marrow stem cells is correlated with in situ localization. Blood 2011; 117(19): 5067-77.
 
[47]  Boxall SA, Jones E. Markers for characterization of bone marrow multipotential stromal cells.Stem Cells Int 2012; 2012: 975871.
 
[48]  Rasini V, Dominici M, Kluba T, Siegel G, Lusenti G, Northoff H, Horwitz EM, Schäfer R. Mesenchymal stromal/stem cells markers in the human bone marrow. Cytotherapy 2013; 15(3):2 92-306.
 
[49]  Spoerri PE, Ludwig HC, Ogawa Y. Amodified silver method for demonstrating developing nervous tissue in culture. ActaAnatomica 1985; 123(1): 64-6.
 
[50]  Lovatt D, Sonnewald U, Waagepetersen HS, Schousboe A, He W, Lin JH, Han X, Takano T, Wang S, Sim FJ, Goldman SA, Nedergaard M.The transcriptome and metabolic gene signature of protoplasmic astrocytes in the adult murine cortex. J Neurosci 2007; 27(45): 12255-66.
 
[51]  Brunet JF, Allaman I, Magistretti PJ, Pellerin L. Glycogen metabolism as a marker of astrocyte differentiation.J Cereb Blood Flow Metab 2010; 30(1): 51-5.
 
[52]  LehtinenMK, Bjornsson CS, Dymecki SM, Gilbertson RJ, Holtzman DM, MonukiES.The Choroid Plexus and Cerebrospinal Fluid: Emerging Roles in Development, Disease, and Therapy. The Journal of Neuroscience 2013; 33(45): 17553-9.
 
[53]  Taran R, Mamidi MK, Singh G, Dutta S, Parhar IS, John JP, Bhonde R, Pal R, Das AK. In vitro and in vivo neurogenic potential of mesenchymal stem cells isolated from different sources. J Biosci 2014; 39: 157-69.