JBiSE  Vol.6 No.4 , April 2013
Neural differentiation of allogenic mixed-cultured rat bone mesenchymal stem cells
ABSTRACT

Many studies showed that bone mesenchymal stem cells (BMSCs) can transdifferentiate to neural cell in vitro. The purpose of this study was to investigate the mixed-culture of allogenic rat BMSCs in vitro, and to perform neural differentiation potential characterization. Rat BMSCs were isolated and cultured by plastic adherence and density gradient centrifugation respectively, and the 3rd passage cells were harvested and mixed-cultured with same concentration. The second passage cells of the mixed-cultured cells were obtained to perform Wright-Gemsa staining for morphological observation, to identify the surface marker of CD29, CD45, CD90 by flow cytometry and induced to differentiate into neural cell, then performed immunocytochemistry of Nestin, NSE and GFAP. Results showed that after Wright-Giemsa staining, the mixed-cultured cells displayed typical spindle-shape, presented layered and whirlpool-like growth; and the mixed-cultured cells were positive for CD29 and CD90, but negative for CD45 by flow cytometry. After induction, the mixed-cultured cells appeared morphological changes of neuron and glial cell, and were positive expression of Nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP). The results demonstrated that the mixed-cultured allogenic rat BMSCs can be induced into neural cell such as neuron and glial cell in vitro.


Cite this paper
Zhou, J. , Yang, H. and Cen, J. (2013) Neural differentiation of allogenic mixed-cultured rat bone mesenchymal stem cells. Journal of Biomedical Science and Engineering, 6, 466-472. doi: 10.4236/jbise.2013.64058.
References
[1]   Lindvall, O., Kokaia, Z. and Martinez-Serrano, A. (2004) Stem cell therapy for human neurodegenerative disorders—How to make it work. Nature Medicine, 10, S42- S50. doi:10.1038/nm1064

[2]   Leipzig, N.D., Wylie, R.G., Kim, H. and Shoichet, M.S. (2011) Differentiation of neural stem cells in three-dimensional growth factor—Immobilized chitosan hydrogel scaffolds. Biomaterials, 32, 57-64. doi:10.1016/j.biomaterials.2010.09.031

[3]   Rak, K., Wasielewski, N.V., Radeloff, A., Völkers, J., Scherzed, A., Jablonka, S., Hagen, R. and Mlynski, R. (2011) Isolation and characterization of neural stem cells from the neonatal rat cochlear nucleus. Cell Tissue Research, 343, 499-508. doi:10.1007/s00441-010-1118-x

[4]   McMahon, S.S., Albermann, S., Rooney, G.E., Shaw, G., Garcia, Y., Sweeney, E., Hynes, J., Dockery, P., O’Brien, T., Windebank, A.J., Allsopp, T.E. and Barry, F.P. (2010) Engraftment, migration and differentiation of neural stem cells in the rat spinal cord following contusion injury. Cytotherapy, 12, 313-325. doi:10.3109/14653241003695018

[5]   Mitrecic, D., Nicaise, C., Gajovic, S. and Pochet, R. (2010) Distribution, differentiation, and survival of intravenously administered neural stem cells in a rat model of amyotrophic lateral sclerosis. Cell Transplant, 19, 537-548. doi:10.3727/096368910X498269

[6]   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, 364-370. doi:10.1002/1097-4547(20000815)61:4<364::AID-JNR2>3.0.CO;2-C

[7]   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. doi:10.1016/j.cellbi.2007.02.006

[8]   Bronzi, D., Bramanti, V., Tomassoni, D., Laureanti, F., Grasso, S., Li Volsi, G. and Avola, R. (2010) Neural markers expression in rat bone marrow mesenchymal stem cell cultures treated with neurosteroids. Neurochemical Research, 35, 2154-2160. doi:10.1007/s11064-010-0283-3

[9]   Tseng, P.Y., Chen, C.J., Sheu, C.C., Yu, C.W. and Huang, Y.S. (2007) Spontaneous differentiation of adult rat marrow stromal cells in a long-term culture. Journal of Veterinary Medical Science, 69, 95-102. doi:10.1292/jvms.69.95

[10]   Zhang, L., Kahn, C.J., Chen, H.Q., Tran, N. and Wang, X. (2008) Effect of uniaxial stretching on rat bone mesenchymal stem cell: Orientation and expressions of collagen types I and III and tenascin-C. Cell Biology International, 32, 344-352. doi:10.1016/j.cellbi.2007.12.018

[11]   Huang, Y., Jia, X., Bai, K., Gong, X. and Fan, Y. (2010) Effect of fluid shear stress on cardiomyogenic differentiation of rat bone marrow mesenchymal stem cells. Archives of Medical Research, 41, 497-505. doi:10.1016/j.arcmed.2010.10.002

[12]   Jendelová, P., Herynek, V., DeCroos, J., Glogarová, K., Andersson, B., Hájek, M. and Syková, E. (2003) Imaging the fate of implanted bone marrow stromal cells labeled with superparamagnetic nanoparticles. Magnetic Resonance in Medicine, 50, 767-776. doi:10.1002/mrm.10585

[13]   Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Keating, A., Dj, P. and Horwitz, E. (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8, 315-317. doi:10.1080/14653240600855905

[14]   Harting, M., Jimenez, F., Pati, S., Baumgartner, J. and Cox Jr., C. (2008) Immunophenotype characterization of rat mesenchymal stromal cells. Cytotherapy, 10, 243-253. doi:10.1080/14653240801950000

[15]   Halfon, S., Abramov, N., Grinblat, B. and Ginis, I. (2011) Markers distinguishing mesenchymal stem cells from fibroblasts are downregulated with passaging. Stem Cells and Development, 20, 53-66. doi:10.1089/scd.2010.0040

[16]   Takemitsu, H., Zhao, D., Yamamoto, I., Harada, Y., Michishita, M. and Arai, T. (2012) Comparison of bone marrow and adipose tissue-derived canine mesenchymal stem cells. BMC Veterinary Research, 31, 150. doi:10.1186/1746-6148-8-150

[17]   Vanderwinden, J.M., Gillard, K., De Laet, M.H., Messam, C.A. and Schiffmann, S.N. (2002) Distribution of the intermediate filament nestin in the muscularis propria of the human gastrointestinal tract. Cell Tissue Research, 309, 261-268. doi:10.1007/s00441-002-0590-3

[18]   Wiese, C., Rolletschek, A., Kania, G., Blyszczuk, P., Tarasov, K.V., Tarasova, Y., Wersto, R.P., Boheler, K.R. and Wobus, A.M. (2004) Nestin expression—A property of multi-lineage progenitor cells? Cellular and Molecular Life Sciences, 61, 2510-2522. doi:10.1007/s00018-004-4144-6

[19]   Pillai, R., Scintu, F., Scorciapino, L., Carta, M., Murru, L., Biggio, G., Cabras, S., Reali, C. and Sogos, V. (2006) Human astrocytes can be induced to differentiate into cells with neuronal phenotype. Experimental Cell Research, 312, 2336-2346. doi:10.1016/j.yexcr.2006.03.031

[20]   Tohill, M., Mantovani, C., Wiberg, M. and Terenghi, G. (2004) Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neuroscience Letters, 362, 200-203. doi:10.1016/j.neulet.2004.03.077

[21]   Timmer, M., Cesnulevicius, K., Winkler, C., Kolb, J., Lipokatic-Takacs, E., Jungnickel, J. and Grothe, C. (2007) Fibroblast growth factor (FGF)-2 and FGF receptor 3 are required for the development of the substantia nigra, and FGF-2 plays a crucial role for the rescue of dopaminergic neurons after 6-hydroxydopamine lesion. Journal of Neuroscience, 27, 459-471. doi:10.1523/JNEUROSCI.4493-06.2007

[22]   Tao, H., Rao, R. and Ma, D.D. (2005) Cytokine-induced stable neuronal differentiation of human bone marrow mesenchymal stem cells in a serum/feeder cell-free condition. Development, Growth & Differentiation, 47, 423- 433. doi:10.1111/j.1440-169X.2005.00810.x

[23]   Justesen, J., Stenderup, K. and Kassem, M.S. (2001) Mesenchymal stem cells. Potential use in cell and gene therapy of bone loss caused by aging and osteoporosis. Ugeskr Laeger, 163, 5491-5495.

 
 
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