Remyelination of Demyelinated Rat Spinal Cord Model by Transplanting Neural Stem Cells

Authors
1 M.Sc. Student, Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Professor, Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
3 Neurosurgeon, Shefa Neuroscience Research Center, Khatam al-Anbia Hospital, Tehran, Iran
Abstract
Objective: Demyelination of CNS axons occurs under pathological conditions such as multiple sclerosis and spinal cord injuries, but can be repaired by cell therapy. Within the CNS remyelination can be achieved by transplantation of neural stem cells (NSCs). NSCs are self-renewing cells that maintain the capacity to differentiate into CNS-specific cell types and can differentiate into the three main neural phenotypes: astroglia, oligodendroglia and neurons. They may also replace or repair diseased CNS tissue. Methods: Bone marrow stromal cells (BMSCs) were aseptically isolated from the tibia and femurs of young adult Sprague Dawley rats. BMSCs were evaluated by fibronectin and CD31 markers. BMSC-derived NSCs were evaluated by nestin and NF-68. An ethidium bromide-induced demyelinated dorsal column lesion was produced in young adult rats. Transplanting NSCs derived-BMSCs into demyelinated lesions after 3 days in adult rat spinal cords was done. Three weeks after transplantation of NSCs, the spinal cords were processed to evaluate remyelination by Luxol fast blue staining. Results: After passage 3, BMSCs were evaluated and the result, showed the percentage of immunoreactive cells to fibronectin (94.7±2.65), however BMSC-derived NSCs expressed nestin (86.15±0.64) and NF-68 (84.55±0.94) which correlated with fibronectin down regulation. Histologically, the lesions showed slightly irregular elongated areas and had an average length of 1336.36±39.43 µm. Transplanted NSCs were capable of eliciting remyelination. Conclusion: These data support the conclusion that transplantation of NSCs results in functional remyelination of a dorsal column lesion and have valuable applications in the treatment of neurodegenerative diseases such as spinal cord injuries.

Keywords


[1]     Bracken MB. Steroids for acute spinal cord injury. Cochrane Database Syst Rev 2002; (3): CD001046.
[2]     Lim PA, Tow AM. Recovery and regeneration after spinal cord injury: a review and summary of recent literature. Ann Acad Med Singapore 2007; 36(1): 49-57.
[3]     Keirstead HS, Nistor G, Bernal G, Totoiu M, Cloutier F, Sharp K, Steward O. Human embryonic stem cell-derived oligodendrocyte progenitor cell transplants remyelinate and restore locomotion after spinal cord injury. J Neurosci 2005; 25(19): 4694-705.
[4]     Lakatos A, Smith PM, Barnett SC, Franklin RJ. Meningeal cells enhance limited CNS re-myelination by transplanted olfactory ensheathing cells. Brain 2002; 126(Pt 3): 598-609.
[5]     Suzuki H, Taguchi T, Tanaka H, Kataoka H, Li Z, Muramatsu K, Gondo T, Kawai S. Neurospheres induced from bone marrow stromal cells are multipotent for differentiation into neuron, astrocyte, and oligodendrocyte phenotypes. Biochem Biophys Res Commun 2004; 322(3): 918-22.
[6]     Sasaki M, Honmou O, Akiyama Y, Uede T, Hashi K, Kocsis JD. Transplantation of an acutely isolated bone marrow fraction repairs demyelinated adult rat spinal cord axons. Glia 2001; 35(1): 26-34.
[7]     Brochner CB, Johansen JS, Larsen LA, Bak M, Mikkelsen HB, Byskov AG, Andersen CY, Mollgard K. YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers. J Histochem Cytochem 2012; 60(3): 188-204.
[8]     Fujita Y, Inokuma D, Abe R, Sasaki M, Nakamura H, Shimizu T, Shimizu H. Conversion from human haematopoietic stem cells to keratinocytes requires keratinocyte secretory factors. Clin Exp Dermatol 2012. [Epub ahead of print]
[9]     Akiyama Y, Radtke C, Kocsis JD. Remyelination of the rat spinal cord by transplantation of identified bone marrow stromal cells. J Neurosci 2002; 22(15): 6623-30.
[10]  Lindvall O, Kokaia Z. Stem cells for the treatment of neurological disorders. Nature 2006; 441(7097): 1094-6.
[11]  Mothe AJ, Tator CH. Transplanted neural stem/progenitor cells generate myelinating oligodendrocytes and Schwann cells in spinal cord demyelination and dysmyelination. Exp Neurol 2008; 213(1): 176-90.
[12]  Kotter MR, Li WW, Zhao C, Franklin RJ. Myelin impairs CNS remyelination by inhibiting oligodendrocyte precursor cell differentiation. J Neurosci 2006; 26(1): 328-32.
[13]  Burrell AM, Handel AE, Ramagopalan SV, Ebers GC, Morahan JM. Epigenetic mechanisms in multiple sclerosis and the major histocompatibility complex (MHC). Discov Med 2011; 11(58): 187-96.
[14]  Vermersch P, Benrabah R, Schmidt N, Zéphir H, Clavelou P, Vongsouthi C, Dubreuil P, Moussy A, Hermine O. Masitinib treatment in patients with progressive multiple sclerosis: a randomized pilot study. BMC Neurol 2012; 12(1): 36.
[15]  Ide C, Kitada M, Chakrabortty S, Taketomi M, Matsumoto N, Kikukawa S, Mizoguchi A, Kawaguchi S, Endoh K, Suzuki Y. Grafting of choroid plexus ependymal cells promotes the growth of regenerating axons in the dorsal funiculus of rat spinal cord: a preliminary report. Exp Neurol 2001; 167(2): 242-51.
[16]  Wehner R, Taubert C, Mende T, Gaebler C, de Andrade AV, Bornhäuser M, Werner C, Tonn T, Schäkel K, Bachmann M, Schmitz M. Engineered extracellular matrix components do not alter the immunomodulatory properties of mesenchymal stromal cells in vitro. J Tissue Eng Regen Med 2012. [Epub ahead of print]
[17]  Izadpanah R, Joswig T, Tsien F, Dufour J, Kirijan JC, Bunnell BA. Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques. Stem Cells Dev 2005; 14(4): 440-51.
[18]  Yang Q, Mu J, Li Q, Li A, Zeng Z, Yang J, Zhang X, Tang J, Xie P. A simple and efficient method for deriving neurospheres from bone marrow stromal cells. Biochem Biophys Res Commun 2008; 372(4): 520-4.
[19]  Lankford KL, Imaizumi T, Honmou O, Kocsis JD. A quantitative morphometric analysis of rat spinal cord remyelination following trans-plantation of allogenic Schwann cells. J Comp Neurol 2002; 443(3): 259-74.
[20]  Wu S, Suzuki Y, Noda T, Bai H, Kitada M, Kataoka K, Nishimura Y, Ide C. Immunohistochemical and electron micro-scopic study of invasion and differentiation in spinal cord lesion of neural stem cells grafted through cerebrospinal fluid in rat. J Neurosci Res 2002; 69(6): 940-5.
[21]  Kumagai G, Okada Y, Yamane J, Nagoshi N, Kitamura K, Mukaino M, Tsuji O, Fujiyoshi K, Katoh H, Okada S, Shibata S, Matsuzaki Y, Toh S, Toyama, Nakamura M, Okano H. Roles of ES cell-derived gliogenic neural stem/progenitor cells in functional recovery after spinal cord injury. PLoS One 2009; 4(11): e7706.
[22]  Steinbrecher A, Weber T, Neuberger T, Mueller AM, Pedré X, Giegerich G, Bogdahn U, Jakob P, Haase A, Faber C. Experimental autoimmune encephalomyelitis in the rat spinal cord: lesion detection with high-resolution MR microscopy at 17.6 T. AJNR Am J Neuroradiol 2005; 26(1): 19-25.