Production of Neural Progenitors from Bone Marrow Mesenchymal Stem Cells


In the brain, there are hundreds of types of specialized neurons and to generate one type of them we need to have neural progenitors for differentiation to specific neuron type. Mesenchymal stem cells (MSCs) are easily isolated, cultured, manipulated ex vivo, showing great potential for therapeutic applications. The adult MSCs have the potential to produce progeny that differentiate into a variety of cell types such as neurons. This fact suggests that MSCs derived neurons are an important cell type and a deep understanding of the molecular characteristics of it would significantly enhance the advancement of cell therapy for neurological disorders. Therefore, in this study, we isolated, identified, and studied neural progenitors by measuring expression levels through neurogenesis pathway of three neural differentiation markers nestin (NES), neurofilament (NF-L), and microtubule association protein (MAP-2) from mouse bone marrow MSCs (mouse bmMSCs) by using butylated hydroxyanisole (BHA) and diethyl sulfoxide (DMSO) as neural inducers agents. The results of immunocytochemistry and Real Time-PCR showed that in contrast to MSCs, neural differentiated cells showed neural progenitor pattern by showing stable increase in NES gene expression through differentiation process with increasing the protein expression through different exposures times, while NF-L gene and protein expression start to increased after 48 h but not replaced the NES expression completely even when its expression passed NES levels. The maturation marker Map-2 expression was low during the duration of differentiation period in protein and gene expression, which prove that these cells are still progenitors and can be redirected into specific type of neurons by further treatments.

Share and Cite:

Mohammad, M. , Yaseen, N. , Al-Joubory, A. , Abdullah, R. , Mahmood, N. , Ahmed, A. and Al-Shammari, A. (2016) Production of Neural Progenitors from Bone Marrow Mesenchymal Stem Cells. Stem Cell Discovery, 6, 1-12. doi: 10.4236/scd.2016.61001.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Nöth, U., Osyczka, A.M., Tuli, R., et al. (2002) Multilineage Mesenchymal Differentiation Potential of Human Trabecular Bone-Derived Cells. Journal of Orthopaedic Research, 20, 1060-1069.
[2] Tuan, R.S., Boland, G. and Tuli, R. (2003) Adult Mesenchymal Stem Cells and Cell-Based Tissue Engineering. Arthritis Research and Therapy, 5, 32-45.
[3] Pittenger, M.F., Mackay, A.M. and Beck, S.C. (1999) Multilineage Potential of Adult Human Mesenchymal Stem Cells. Science, 284, 143-147.
[4] Al-Shammari, A.M., Al-Joboury, A.A. and Yaseen, N.Y. (2012) Isolation and Culture of Neuronal Stem Cells Which Directed into Purkinje Cells to Be Used for Brain Damage and Epilepsy Treatment in Mice. Molecular Therapy, 20, 201-202.
[5] Lee, K.D., Kuo, T. and Whang-Peng J. (2004) In Vitro Hepatic Differentiation of Human Mesenchymal Stem Cells. Hepatology, 40, 1275-1284.
[6] Alshammari, A.M., Salman, M.I., Umran, M.A., et al. (2013) Chondrogenic Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells, First Successes in Iraq. XX International Congress of Mediterranean Federation of Health and Production of Ruminants, Egypt, 19-22 February 2013.
[7] Moon, H.E., Seung, H.Y., Yong, S.H., et al. (2013) Mitochondrial Dysfunction of Immortalized Human Adipose Tissue-Derived Mesenchymal Stromal Cells from Patients with Parkinson’s Disease. Experimental Neurobiology, 22, 283-300.
[8] Sadan, O., Shemesh, N., Barzilay, R., et al. (2012) Mesenchymal Stem Cells Induced to Secrete Neurotrophic Factors Attenuate Quinolinic Acid Toxicity: A Potential Therapy for Huntington’s Disease. Experimental Neurology, 234, 417-427.
[9] Mazzini, L., Mareschi, K., Ferrero, I., et al. (2012) Mesenchymal Stromal Cell Transplantation in Amyotrophic Lateral Sclerosis: A Long-Term Safety Study. Cytotherapy, 14, 56-60.
[10] Karina, O.G., Felipe, L.M., Priscila, K.M., et al. (2014) Therapeutic Effects of the Transplantation of VEGF over Expressing Bone Marrow Mesenchymal Stem Cells in the Hippocampus of Murine Model of Alzheimer’s Disease. Frontiers in Aging Neuroscience, 6, 30.
[11] Sagara, J. and Makino, N. (2008) Glutathione Induces Neuronal Differentiation in Rat Bone Marrow Stromal Cells. Neurochemical Research, 33, 16-21.
[12] Abouelfetouh, A., Kondoh, T., Ehara, K. and Kohmura, E. (2004) Morphological Differentiation of Bone Marrow Stromal Cells into Neuron-Like Cells after Co-Culture with Hippocampal Slice. Brain Research, 1029, 114-119.
[13] Qian, L. and Saltzman, W.M. (2004) Improving the Expansion and Neuronal Differentiation of Mesenchymal Stem Cells through Culture Surface Modification. Biomaterials, 25, 1331-1337.
[14] Lee, J.H., Yu, H.-S., Lee, G.-S., Ji, A., Hyun, J.K. and Kim, H.-W. (2011) Collagen Gel Three-Dimensional Matrices Combined with Adhesive Proteins Stimulate Neuronal Differentiation of Mesenchymal Stem Cells. Journal of the Royal Society Interface, 8, 998-1010.
[15] Nichols, J.E., Jean, A.N., Douglas, D.W., et al. (2013) Neurogenic and Neuro-Protective Potential of a Novel Subpopulation of Peripheral Blood-Derived CD133+ ABCG2+CXCR4+ Mesenchymal Stem Cells: Development of Autologous Cell Based Therapeutics for Traumatic Brain Injury. Stem Cell Research & Therapy, 4, 3.
[16] Feng, Z. and Gao, F. (2012) Stem Cell Challenges in the Treatment of Neurodegenerative Disease. CNS Neuroscience & Therapeutics, 18, 142-148.
[17] Huang, H., Lin, C. and Paul, S. (2010) Cell Therapy from Bench to Bedside Translation in CNS Neurorestoratology Era. Cell Medicine, 1, 15-46.
[18] Zhao, L., Lin, Y.D., Ma, J., Sun, Y.Y., Zeng, S.J., Zhang, X.W. and Zuo, M.X. (2007) Culture and Neural Differentiation of Rat Bone Marrow Mesenchymal Stem Cells in Vitro. Cell Biology International, 31, 916-923.
[19] Cui, P., He, X.H., Pu, Y.B., Zhang, W.X., Zhang, P., Li, C.L., Guan, W.J., Li, X.C. and Ma, Y.H. (2014) Biological Characterization and Pluripotent Identification of Sheep Dermis-Derived Mesenchymal Stem/Progenitor Cells. BioMed Research International, 2014, Article ID: 786234.
[20] Zhao, G.G., Ji, H.J., Wang, S.H., Gu, B., Song, X.L., Zhang, J.R., Liu, Y.K., Chen, L.B. and Zhang, M. (2014) Cell Surface Proteomics Analysis Indicates a Neural Lineage Bias of Rat Bone Marrow Mesenchymal Stromal Cells. BioMed Research International, 2014, Article ID: 479269.
[21] Mehranjani, S.M. and Chian, F.M. (2014) Cysteine: A Novel Neural Inducer for Rat Bone Marrow Mesenchymal Stem Cells. Cell Journal, 16, 195-202.
[22] Kim, E.Y., Lee, K.-B., Yu, J., et al. (2014) Neuronal Cell Differentiation of Mesenchymal Stem Cells Originating from Canine Amniotic Fluid. Human Cell, 27, 51-58.
[23] Birbrair, A., Wang, Z.M., Messi, M.L., Enikolopov, G.N. and Delbono, O. (2011) Nestin-GFP Transgene Reveals Neural Precursor Cells in Adult Skeletal Muscle. PLoS ONE, 6, e16816.
[24] Leonard, D.B., Gorham, J.D., Cole, P., Greene, L.A. and Ziff, E.B. (1988) A Nerve Growth Factor-Regulated Messenger RNA Encodes a New Intermediate Filament Protein. The Journal of Cell Biology, 106, 181-193.
[25] Wieske, L., Witteveen, E., Petzold, A., Verhamme, C., Schultz, M.J., van Schaik, I.N. and Horn, J. (2014) Neurofilaments as a Plasma Biomarker for ICU-Acquired Weakness: An Observational Pilot Study. Critical Care, 18, R18.
[26] Caceres, A., Banker, G.A. and Binder, L. (1986) Immunocytochemical Localization of Tubulin and Microtubule-Associated Protein 2 during the Development of Hippocampal Neurons in Culture. The Journal of Neuroscience, 6, 714-722.
[27] Fischer, I., Shea, T.B., Sapirstein, V.S. and Kosik, K.S. (1986) Expression and Distribution of Microtubule-Associated Protein 2 (MAP2) in Neuroblastoma and Primary Neuronal Cells. Developmental Brain Research, 25, 99-109.
[28] Freshney, R.I. (2005) Culture of Animal Cells: A Manual for Basic Technique. Fifth Edition, Wiley, New York.
[29] Woodbury, D., Schwarz, E.J., Prockop, D.J. and Black, I.B. (2000) Adult Rat and Human Bone Marrow Stromal Cells Differentiate into Neurons. Journal of Neuroscience Research, 61, 364-367.<364::AID-JNR2>3.0.CO;2-C
[30] Vandesompele, J., De Preter, K., Pattyn, F., Poppe, B., Van Roy, N., De Paepe, A. and Speleman, F. (2002) Accurate Normalization of Real-Time Quantitative RT-PCR Data by Geometric Averaging of Multiple Internal Control Genes. Genome biology, 3, RESEARCH0034.
[31] Livak, K.J. and Schmittgen, T.D. (2001) Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-ΔΔCT Method. Methods, 25, 402-408.
[32] Schmittgen, T.D., Livak, K.J. (2008) Analyzing Real-Time PCR Data by the Comparative CT Method. Nature Protocols, 3, 1101-1108.
[33] Maniatis, T., Freitsch, E. and Sambrook, J. (1989) Gel Electrophoresis of DNA. In: Sambrook, J., Fritsch, E.F. and Maniatis, T., Eds., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor.
[34] Michalczyk, K. and Ziman, M. (2005) Nestin Structure and Predicted Function in Cellular Cytoskeletal Organisation. Histology and Histopathology, 20, 665-671.
[35] Woodbury, D., Reynolds, K. and Black, I.B. (2002) Adult Bone Marrow Stromal Stem Cells Express Germ Line, Ectodermal, Endodermal, and Mesodermal Genes Prior to Neurogenesis. Journal of Neuroscience Research, 69, 908-917.
[36] Harvey, B.S. (2013) Clinical Neuropathology Practice Guide 5-2013: Markers of Neuronal Maturation. Clinical Neuropathology, 32, 340-369.
[37] Delcroix, G.J.-R., Jacquart, M., Lemaire, L., Sindji, L., Franconi, F., Le Jeune, J.-J. and Montero-Menei, C.N. (2009) Mesenchymal and Neural Stem Cells Labeled with HEDP-Coated SPIO Nanoparticles: In Vitro Characterization and Migration Potential in Rat Brain. Brain Research, 1255, 18-31.
[38] Sanchez-Ramos, J., Song, S., Cardozo-Pelaez, F., et al. (2000) Adult Bone Marrow Stromal Cells Differentiate into Neural Cells in Vitro. Experimental Neurology, 164, 247-256.
[39] Zhang, H.B., Huang, Z.Y., Xu, Y.M. and Zhang, S.M. (2006) Differentiation and Neurological Benefit of the Mesenchymal Stem Cells Transplanted into the Rat Brain Following Intracerebral Hemorrhage. Neurological Research, 28, 104-112.
[40] Kim, B.J., Seo, J.H., Bubien, J.K. and Oh, Y.S. (2002) Differentiation of Adult Bone Marrow Stem Cells into Neuroprogenitor Cells in Vitro. Neuroreport, 13, 1185-1188.
[41] Hung, S.C., Chen, N.J., Hsieh, S.L., Li, H., Ma, H.-L. and Lo, W.-H. (2002) Isolation and Characterization of Size-Sieved Stem Cells from Human Bone Marrow. Stem Cells, 20, 249-258.
[42] Iida, K. and Nishimura, I. (2002) Gene Expression Profiling by DNA Microarray Technology. Critical Reviews in Oral Biology & Medicine, 13, 35-50.
[43] Jiang, Y., Jahagirdar, B.N., Reinhardt, R.L., et al. (2002) Pluripotency of Mesenchymal Stem Cells Derived from Adult Marrow. Nature, 418, 41-49.
[44] Egusa, H., Schweizer, F.E., Wang, C.C., Matsuka, Y. and Nishimura, I. (2005) Neuronal Differentiation of Bone Marrow-Derived Stromal Stem Cells Involves Suppression of Discordant Phenotypes through Gene Silencing. The Journal of Biological Chemistry, 280, 23691-23697.
[45] Darabi, S., Tiraihi, T., Delshad, A. and Sadeghizadeh, M. (2013) A New Multistep Induction Protocol for the Transdifferentiation of Bone Marrow Stromal Stem Cells into GABAergic Neuron-Like Cells. Iranian Biomedical Journal, 17, 8-14.
[46] Lalonde, R. and Strazielle, C. (2003) Neurobehavioral Characteristics of Mice with Modified Intermediate Filament Genes. Reviews in the Neurosciences, 14, 369-385.
[47] Shafit-Zagarado, B., Rockwood, J., Davies, P., Kress, Y. and Lee, S.C. (2000) Novel Microtubule-Associated Protein-2 Isoform Is Expressed Early in Human Oligodendrocyte Maturation. Glia, 29, 233-245.<233::AID-GLIA5>3.0.CO;2-U
[48] Mohammad-Gharibani, P., Tiraihi, T. and Arabkheradmand, J. (2009) In Vitro Transdifferentiation of Bone Marrow Stromal Cells into GABAergic-Like Neurons. Iranian Biomedical Journal, 13, 137-143.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.