Ex vivo differentiation of human bone marrow-derived stem cells into neuronal cell-like lineages

Abstract

Ex vivo differentiation of human bone marrow-derived stem cells into neuronal cell-like lineages Adeeb Al-Zoubi,1,2 Feras Altwal,3 Farah Khalifeh,2 Jamil Hermas,4 Ziad Al-Zoubi,5 Emad Jafar,5 Mohammed El-Khateeb,6,7 1Department of Surgery, University of Illinois College of Medicine at Peoria, Peoria, IL, USA; 2Stem Cells of Arabia, Amman, Jordan; 3Department of Neuroscience, School of Graduate and Postdoctoral Studies, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA; 4Stem Cell Division, Al-Yamama Company, 5Jordan Orthopedic and Spinal Center, 6National Center for Diabetes, Endocrinology and Genetics, 7Department of Pathology, Faculty of Medicine, University of Jordan, Amman, Jordan Background: Methods to obtain safe and practical populations of stem cells (SCs) at a clinical grade that are able to differentiate into neuronal cell lineages are yet to be developed. In a previous study, we showed that mouse bone marrow-derived SCs (BM-SCs) differentiated into neuronal cell-like lineages when put in a neuronal-like environment, which is a special media supplemented with the necessary growth factors needed for the differentiation of SCs into neuronal cell-like lineages. Aim: In this study, we aim to assess the potentials of adult human CD34+ and CD133+ SCs to differentiate into neuronal cell-like lineages ex vivo when placed in a neuronal-like microenvironment. Methods: The neuronal-like microenvironment was created by culturing cells in nonhematopoietic expansion media (NHEM) supplemented with growth factors that favor differentiation into neuronal cell lineages (low-affinity nerve growth factor [LNGF], mouse spinal cord extract [mSpE], or both). Cultured cells were assessed for neuronal differentiation by cell morphologies and by expression of GFAP. Results: Our results show that culturing unpurified human BM-derived mononuclear cells (hBM-MNCs) in NHEM+LNGF+mSpE did not lead to neuronal differentiation. In contrast, culturing of purified CD34+ hBM-SCs in NHEM+LNGF+mSpE favored their differentiation into astrocyte-like cells, whereas culturing of purified CD133+ hBM-SCs in the same media favored their differentiation into neuronal-like cells. Interestingly, coculturing of CD34+ and CD133+ hBM-SCs in the same media enhanced the differentiation into astrocyte-like cells and neuronal-like cells, in addition to oligodendrocyte-like cells. Conclusion: These results suggest that a mixture of purified CD34+ and CD133+ cells may enhance the differentiation into neuronal cell-like lineages and give broader neuronal cell lineages than when each of these cell types is cultured alone. This method opens the window for the utilization of specific populations of hBM-SCs to be delivered in a purified form for the potential treatment of neurodegenerative diseases in the future. Keywords: cell therapy, neurodegenerative disease, neurons, astrocyes, CD34+, CD133+, MACS