Neurogenic Differentiation of Bone Marrow-derived Mesenchymal Stem Cells Using Neural Induction Medium: A Morphological and Histochemical Study
American Journal of Bioscience and Bioengineering
Volume 3, Issue 4-1, July 2015, Pages: 43-50
Received: Jun. 26, 2015; Accepted: Jun. 27, 2015; Published: Jul. 6, 2015
Views 2654      Downloads 71
Authors
Ahmedy E., Clinical pathology department, faculty of medicine, El Menoufia universit, Menofia, Egypt
Kandel S., Clinical pathology department, faculty of medicine, El Menoufia universit, Menofia, Egypt
Gabr H., Clinical pathology department, faculty of medicine, Kasr El Eini University, Cairo, Egypt
Rizk S., Clinical pathology department, faculty of medicine, Kasr El Eini University, Cairo, Egypt
Khalifa . K, Clinical pathology department, faculty of medicine, El Menoufia universit, Menofia, Egypt
Kamal S., Clinical pathology department, faculty of medicine, El Menoufia universit, Menofia, Egypt
Article Tools
Follow on us
Abstract
Background: Mesenchymal stem cells (MSCs) are multipotent adult stem cells present in all tissues. They are present in bone marrow, and can differentiate in vitro into neurons, glial cells and myofibroblasts . MSCs have been proposed as sources of stem cells for regeneration of the CNS. Thus, one of the goals of regenerative medicine is to regenerate damaged brain tissue and spinal cord by harnessing the power of stem cells to initiate neurogenesis in damaged areas of the brain. Objective: The aim of this work is to study in-vitro induced neurogenesis using MSCs as model of stem cells. Methodology : Bone marrow-MSCs were isolated, expanded and passaged. MSCs were identified using morphology and flowcytometric analysis. Co-expression of Oct ¾ was done. MSCs were induced to neural lineage using Neural Induction Media (NIM) : a cocktail of retinoic acid dissolved in DEMSO, recombinant human Fibroblast Growth Factor (FGF) basic, recombinant human Epidermal Growth Factor (EGF) and Insulin-like Growth Factor I (IGF-I) . Neural induction was verified morphologically, and immunologically using GFAP positivity and nestin expression. Results: BM-MSCs express CD44 and OCT ¾ which decrease with age. MSCs induced with NIM show morphological changes consistent with neurogenesis, positive GFAP and nestin expression as compared to the uninduced cells. Conclusion: MSCs isolated from bone marrow aspirate and can be differentiated into GFAP positive neural cells.
Keywords
Mesenchymal Stem Cells, GFAP, Retinoic Acid, Neurogenesis
To cite this article
Ahmedy E., Kandel S., Gabr H., Rizk S., Khalifa . K, Kamal S., Neurogenic Differentiation of Bone Marrow-derived Mesenchymal Stem Cells Using Neural Induction Medium: A Morphological and Histochemical Study, American Journal of Bioscience and Bioengineering. Special Issue: Stem Cells for Neuro-Regeneration: Where Do We Stand. Vol. 3, No. 4-1, 2015, pp. 43-50. doi: 10.11648/j.bio.s.2015030401.17
References
[1]
Becker AJ, McCulloch EA and Till JE (1963),: "Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells". Nature197: 452–4.
[2]
Siminovitch L, McCulloch EA and Till JE (1963),: "The distribution of colony-forming cells among spleen colonies". Journal of Cellular and Comparative Physiology62: 327–36.
[3]
Tuch BE (2006): "Stem cells—a clinical update". Australian Family Physician35 (9): 719–21.
[4]
Malgieri A, Kantzari E, Patrizi MP & Gambardella S (2010): Bone marrow and umbilical cord blood human mesenchymal stem cells:state of the art. Int J Clin Exp Med; 3(4):248-69.
[5]
Takashima, Y., Era, T., Nakao, K., Kondo, S., Kasuga, M., Smith, A.G., and Nishikawa, S. (2007): Neuroepithelialcells supply an initial transient wave of MSC differentiation. Cell 129, 1377–1388.
[6]
Paspala, S; Murthy, T; Mahaboob, V; Mahaboob, V and Habeeb, M (2011): "Pluripotent stem cells - A review of the current status in neural regeneration". Neurology India 59 (4): 558-65.
[7]
Alenzi, F and Bahkali, A (2011):"Stem cells: Biology and clinical potential". African Journal of Biotechnology 10 (86): 19929-40.
[8]
Ratajczak M.Z..Zuba-Surma EK ,Paczkowska E, Kucia M, Nowack P.(2011): Stem cell for neural regeneration-a potential application of very small embryonic-like stem cell.Journal of physiology and pharmacology., 62, 1, 3-12.
[9]
Farin A, Liu CY, Langmoen IA and Apuzzo ML (2009): Biological restoration of central nervous system architecture and function: part 3-stem cell- and cell-based applications andrealities in the biological management of central nervoussystem disorders: traumatic, vascular, and epilepsy disorders. Neurosurgery; 65: 831-859.
[10]
Galindo Layla T., Filippo Thais R. M., Patricia Semedo, Ariza Carolina B., Moreira Caroline M. et al.,(2011): Mesenchymal Stem Cell Therapy Modulates the Inflammatory Response in Experimental Traumatic Brain Injury* Neurol Res Int. 2011; 2011: 564089. doi: 10.1155/2011/564089
[11]
Dexheimer V, Mueller S, Braatz F and Richter W (2011) Reduced Reactivation from Dormancy but Maintained Lineage Choice of Human Mesenchymal StemCells with Donor Age. PLoS ONE 6(8): e22980.
[12]
Ishizaka R, Hayashi Y, Iohara K, Sugiyama M, Murakami M, Yamamoto T, Fukuta O, and Nakashima M.(2013): Stimulation of angiogenesis, neurogenesis and regeneration by side population cells from dental pulp. Biomaterials.34(8):1888-97. doi: 10.1016/j.biomaterials.2012.10.045. Epub 2012 Dec 11.
[13]
Fu L, Zhu L, Huang Y, Lee T, Forman S, and Shih C . (2008): Derivation of Neural Stem Cells from Mesenchymal Stem Cells: Evidence for a Bipotential Stem Cell Population Stem Cells Dev. 17(6): 1109–1121.
[14]
Khoo Melissa L. M, Tao Helen, Meedeniya Adrian C. B., Mackay-Sim Alan, and Ma David D. F.(2011):Transplantation of Neuronal-Primed Human Bone Marrow Mesenchymal Stem Cells in Hemiparkinsonian Rodents PLoS One. 6(5): e19025.
[15]
Maijenburg MW, Kleijer M, Vermeul K, Mul E, van Alphen FP,van der Schoot CE, and Voermans C.(2011): The composition of the mesenchymal stromalcell compartment in human bone marrow changes during development and aging.Haematologica. 96:xxxdoi:10.3324/haematol.2011.047753
[16]
Galbraith PR. (1974) :Studies on control of granulopoiesis in man II. Influence of circulating neutrophil count on release of labelled bone marrow cells Can Med Assoc J. November 2; 111(9): 919–923. PMCID: PMC1955909.
[17]
Lo T, Ho JH, Yang MH & Lee OK (2010): Glucose reduction prevents replicative senescence and increases mitochondrial respiration in human mesenchymal stem cells. Cell Transplant Nov 5.
[18]
Orciani M, Mariggio MA, Morabito C, Di BG, Di PR.(2010): Functional characterization of calcium-signaling pathways of human skin-derived mesenchymal stem cells. Skin Pharmacol Physiol.23(3):124–32.
[19]
Hasebe Y, Hasegawa S, Hashimoto N, et al.(2011): Analysis of cell characterization using cell surface markers in the dermis. J Dermatol Sci.62(2):98–106.
[20]
Zvaifler NJ, Marinova-Mutafchieva L, Adams G, et al.(2000): Mesenchymal precursor cells in the blood of normal individuals. Arthritis Res.2(6):477–88.
[21]
LIN Yu-mei, ZHANG Gui-zhen, LENG Zong-xiang, LU Zhen-xia, BU Li-sha, GAO Shen, YANG Shao-juan.(2006): Study on the bone marrow mesenchymal stem cells induced drug resistance in the U937 cells and its mechanism. Chinese Medical Journal. Vol. 119 No. 11 : 905-910.
[22]
Sackstein R (2009). "Glycosyltransferase-programmed stereosubstitution (GPS) to create HCELL: engineering a roadmap for cell migration". Immunol. Rev.230 (1): 51–74.
[23]
Alves CS, Yakovlev S, Medved L, Konstantopoulos K. (2009): "Biomolecular characterization of CD44-fibrin(ogen) binding: distinct molecular requirements mediate binding of standard and variant isoforms of CD44 to immobilized fibrin(ogen)". J Biol Chem284 (2): 1177–89.
[24]
Ishii M, Koike C, Igarashi A et al. (2005): Molecular markers distinguish bone marrow mesenchymal stem cells from fibroblasts. Biochem Biophys Res Commun.332:297–303.
[25]
Jones E. and McGonagle D. (2008): Human bone marrow mesenchymal stem cells in vivo.Rheumatology.47;126–131
[26]
Miura M, Gronthos S, Zhao M, et al. (2003): Stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA. 100(10):5807–12.
[27]
Kim JH, Jee MK, Lee SY, et al. (2009): Regulation of adipose tissue stromal cells behaviors by endogenic Oct4 expression control". PLoS ONE4 (9): e7166.
[28]
Stolzing A, Jones E, McGonagle D and Scutt A. (2008): Age-related changes in human bone marrow-derived mesenchymal stem cells: consequences for cell therapies. Mech Ageing Dev. 129:163–173.
[29]
Payne KA, Didiano DM and Chu CR (2010): Donor sex and age influence the chondrogenic potential of human femoral bone marrow stem cells. Osteoarthritis Cartilage. 18(5):705-13.
[30]
Khoo ML, Shen B, Tao H and Ma DD. (2008): Long-term serial passage and neuronal differentiation capability of human bone marrow mesenchymal stem cells. Stem Cells Dev. 17:883–896.
[31]
Bertani N, Malatesta P, Volpi G, Sonego P and Perris R. (2005): Neurogenic potential of human mesenchymal stem cells revisited: analysis by immunostaining, time-lapse video and microarray. J Cell Sci. 118:3925–3936.
[32]
Maden M (2007) Retinoic acid in the development, regeneration and maintenance of the nervous system.Nat Rev Neurosci. 8(10):755-65.
[33]
Montiel-Eulefi E, Nery AA, Rodrigues LC, Sánchez R, Romero F, Ulrich H. (2012): Neural differentiation of rat aorta pericyte cells. Cytometry A. 81(1):65-71.
[34]
Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman TB, Saporta S, Janssen W, Patel N, et al. (2000): Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol. 164:247–256.
[35]
Reyes,M., Lund, T., Lenvik, T., Aguiar, D., Koodie, L., and Verfaillie, C.M. (2001). Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98, 2615–2625.
[36]
Sanchez-Ramos JR (2002): Neural cells derived from adult bone marrow and umbilical cord blood. J Neurosci Res; 69(6):880-93.
[37]
Kohyama J, Abe H, Shimazaki T, Koizumi A, Nakashima K, Gojo S, Taga T, Okano H, Hata J & Umezawa A (2001): Brain from bone: Efficient “meta-differentiation” of marrow stroma-derived mature osteoblasts to neurons with Noggin or a demethylating agent. Differentiation; 68:235–44.
ADDRESS
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
U.S.A.
Tel: (001)347-983-5186