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Year : 2013  |  Volume : 26  |  Issue : 2  |  Page : 71-77

Methods and applications for mesenchymal stem cells

1 Department of Clinical Pathology, Faculty of Medicine, El Menoufia University, Menufia, Egypt
2 Department of Clinical Pathology, Faculty of Medicine, Kasr El Eini University, Cairo, Egypt

Correspondence Address:
Eman A Ahmedy
Department of Clinical Pathology, Faculty of medicine, El Menoufia University, Yassin Abd El Ghaffar street, Shebin El Kom, 32511 El Menoufia
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/1110-2098.126094

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Objective The aim of this work is to study neurogenesis using mesenchymal stem cells (MSCs) as a model of stem cells and then follow them as they form neurons. Background MSCs are multipotent adult stem cells present in all tissues. They are present in the 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 central nervous system. Thus, one of the goals of regenerative medicine is to ameliorate irreversible destruction of brain tissue and spinal cord by harnessing the power of stem cells to initiate neurogenesis in damaged areas of the brain. Materials and methods MSCs were cultured from bone marrow aspirate and detected morphologically and by flow cytometric analysis of surface markers CD44 and Oct3/4, then differentiated into neural cells using neural induction media, which consisted of a cocktail of retinoic acid dissolved in DEMSO, recombinant human basic fibroblast growth factor, recombinant human epidermal growth factor, and insulin-like growth factor I, and detected by glial fibrillary acidic protein (GFAP). Results The results of this study showed that MSCs could be isolated from the bone marrow and assumed the typical fibroblastoid morphology and reached 80-90% confluence at about 9 days. They expressed CD44 with a mean ± SD of 81.54 ± 11.58 and CD Oct3/4 with a mean ± SD of 56.12 ± 17.37. MSCs showed positive expression for double expression of CD44-OCT3/4, with a mean ± SD of 54.03 ± 17.42. A highly significant statistical correlation (P < 0.001) was found between age and double expression of CD44-OCT3/4. No statistically significant correlation (P > 0.05) was found between MNCs and double expression of CD44-OCT3/4. MSCs induced with neural induction media show morphological changes consistent with neurogenesis as compared with the symmetric morphologies of the uninduced cells, as shown by an inverted microscope. Induced cells showed positive staining with GFAP whereas uninduced cells showed negative staining. Conclusion MSCs can be isolated successfully from bone marrow aspirate and can be differentiated into GFAP-positive neural cells.

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