Neuroprotective effects of bone marrow stromal cells in cocultures of fetal dopaminergic neuronal cultures

Shijie Song, MD, Amanda Rowe, BS, Kunyu Li, BS, Vasyl Sava, PhD, Juan Sanchez-Ramos, PhD, MD


Objective: The primary objective was to compare the neuroprotection conferred by diffusible factors to those mediated by direct cell-cell contacts in cocultures of bone marrow stromal cells (BMSC) and dopaminergic (DA) neurons treated with methyl-phenyl-pyridinium (MPP+).

Methods: Fetal midbrain (MB) cell cultures of DA neurons were cocultured directly with green fluorescent protein (GFP+)-expressing BMSC or in bilayer cultures separated by a 0.4-µm pore semipermeable membrane. Endpoints of toxicity included survival of tyrosine hydroxylase (TH+) immunoreactive cells, lengths of the TH+ neurites, and [3H]-DA uptake. Neurotrophic factors released into media were measured. A number of cells coexpressing GFP+ and neuronal markers were counted to assess possible fusion with or transdifferentiation into DA neurons.

Results: DA neurons in cocultures (mixed and bilayer) sustained significantly less damage compared to the effects of MPP+ on MB monolayer cultures. Neurite length and [3H]-DA uptake were significantly greater in bilayer cultures than in monolayer MB cell cultures. Mixed cocultures (with MB and GFP+ BMSC in direct contact) were also protected against MPP+. Levels of glial cell-derived neurotrophic factor were found to be elevated in the bilayer culture media. There were no TH+ cells that coexpressed GFP, but 7-10 percent of neurons coexpressed GFP and neuron-specific nuclear antigen suggesting possible fusion of GFP+ BMSC with non-TH+ neurons.

Conclusion: Neuroprotection was observed to an equal extent in both mixed cultures and bilayer cultures in which MB and BMSC cultures were separated by a membrane. Therefore, diffusible factors elaborated by BMSC are sufficient for mitigating neurotoxicity to DA neurons.

Keywords: Mesenchymal stem cells, Methyl-phenyl-pyridinium, Growth factors, Cell fusion


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Chen J, Li Y, Chopp M: Intracerebral transplantation of bone marrow with BDNF after MCAo in rat. Neuropharmacology. 2000; 39: 711-716.

Chen J, Li Y, Wang L, et al.: Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke. 2001; 32: 1005-1011.

Li Y, Chen J, Chopp M: Adult bone marrow transplantation after stroke in adult rats. Cell Transplant. 2001; 10: 31-40.

Li Y, Chen J, Wang L, et al.: Treatment of stroke in rat with intracarotid administration of marrow stromal cells. Neurology. 2001; 56: 1666-1672.

Li Y, Chen J, Wang L, et al.: Intracerebral transplantation of bone marrow stromal cells in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson’s disease. Neurosci Lett. 2001; 316: 67-70.

Li Y, Chopp M, Chen J, et al.: Intrastriatal transplantation of bone marrow nonhematopoietic cells improves functional recovery after stroke in adult mice. J Cereb Blood Flow Metab. 2000; 20: 1311-1319.

Zhao LR, Duan WM, Reyes M, et al.: Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol. 2002; 174: 11-20.

Alvarez-Dolado M, Pardal R, Garcia-Verdugo JM, et al.: Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature. 2003; 425: 968-973.

Terada N, Hamazaki T, Oka M, et al.: Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion. Nature. 2002; 416: 542-545.

Weimann JM, Charlton CA, Brazelton TR, et al.: Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains. Proc Natl Acad Sci USA. 2003; 100: 2088-2093.

Weimann JM, Johansson CB, Trejo A, et al.: Stable reprogrammed heterokaryons form spontaneously in Purkinje neurons after bone marrow transplant. Nat Cell Biol. 2003; 5: 959-966.

Ying Q, Nichols J, Evans EP, et al.: Changing potency by spontaneous fusion. Nature. 2002; 416: 545-548.

Kokovay E, Cunningham LA: Bone marrow-derived microglia contribute to the neuroinflammatory response and express iNOS in the MPTP mouse model of Parkinson’s disease. Neurobiol Dis. 2005; 19: 471-478.

Chen Q, Long Y, Yuan X, et al.: Protective effects of bone marrow stromal cell transplantation in injured rodent brain: Synthesis of neurotrophic factors. J Neurosci Res. 2005; 80: 611-619.

Chen X, Katakowski M, Li Y, et al.: Human bone marrow stromal cell cultures conditioned by traumatic brain tissue extracts: Growth factor production. J Neurosci Res. 2002; 69: 687-691.

Chen X, Li Y, Wang L, et al.: Ischemic rat brain extracts induce human marrow stromal cell growth factor production. Neuropathology. 2002; 22: 275-279.

Garcia R, Aguiar J, Alberti E, et al.: Bone marrow stromal cells produce nerve growth factor and glial cell line-derived neurotrophic factors. Biochem Biophys Res Commun. 2004; 316: 753-754.

Gu Y, Wang J, Ding F, et al.: Neurotrophic actions of bone marrow stromal cells on primary culture of dorsal root ganglion tissues and neurons. J Mol Neurosci. 2010; 40: 332-341.

Kamei N, Tanaka N, Oishi Y, et al.: Bone marrow stromal cells promoting corticospinal axon growth through the release of humoral factors in organotypic cocultures in neonatal rats. J Neurosurg Spine. 2007; 6: 412-419.

Song S, Kamath S, Vesely DL, et al. Expression of brain natriuretic peptide by human bone marrow stromal cells. Paper presented at the Society for Neuroscience Abstract, 2002; No. 824.3.

Song S, Sanchez-Ramos J: Preparation of neural progenitors from bone marrow and umbilical cord blood. In Zigova T, Sanberg PR, Sanchez-Ramos J (eds.): Protocols for Neural Stem Cell Methods. Vol. 198. Totowa, NJ: Human Press, 2002: 79-88.

Michel PP, Dandapani BK, Sanchez-Ramos J, et al.: Toxic effects of potential environmental neurotoxins related to 1-methyl-4-phenylpyridinium on cultured rat dopaminergic neurons. J Pharmacol Exp Therap. 1989; 248: 842-850.

Sanchez-Ramos JR, Song S, Facca A, et al.: Transgenic murine dopaminergic neurons expressing human Cu/Zn superoxide dismutase exhibit increased density in culture, but no resistance to methylphenylpyridinium-induced degeneration. J Neurochem. 1997; 68: 58-67.

Hess DC, Hill WD, Martin-Studdard A, et al.: Bone marrow as a source of endothelial cells and NeuN-expressing cells after stroke. Stroke. 2002; 33: 1362-1368.

Sawada H, Ibi M, Kihara T, et al.: Neuroprotective mechanism of glial cell line-derived neurotrophic factor in mesencephalic neurons. J Neurochem. 2000; 74: 1175-1184.

Sharma HS: Post-traumatic application of brain-derived neurotrophic factor and glia-derived neurotrophic factor on the rat spinal cord enhances neuroprotection and improves motor function. Acta Neurochir Suppl. 2006; 96: 329-334.

Zhang L, Fletcher-Turner A, Marchionni MA, et al.: Neurotrophic and neuroprotective effects of the neuregulin glial growth factor-2 on dopaminergic neurons in rat primary midbrain cultures. J Neurochem. 2004; 91: 1358-1368.

Song S, Kamath S, Mosquera D, et al.: Expression of brain natriuretic peptide by human bone marrow stromal cells. Exp Neurol. 2004; 185: 191-197.

Airaksinen MS, Saarma M: The GDNF family: Signalling, biological functions and therapeutic value. Nat Rev Neurosci. 2002; 3: 383-394.

Nutt JG, Burchiel KJ, Comella CL, et al. Randomized, doubleblind trial of glial cell line-derived neurotrophic factor (GDNF) in PD. Neurology. 2003; 60: 69-73.

Gill SS, Patel NK, Hotton GR, et al.: Direct brain infusion of glial cell line-derived neurotrophic factor in Parkinson disease. Nat Med. 2003; 9: 589-595.

Sanchez-Ramos J, Song S, Cardozo-Pelaez F, et al.: Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol. 2000; 164: 247-256.

Blau HM: A twist of fate. Nature. 2002; 419: 437.

Jiang Y, Jahagirdar BN, Reinhardt RL, et al.: Pluripotency of mesenchymal stem cells derived from adult marrow. Nature. 2002; 418: 41-49.

Kohyama J, Abe H, Shimazaki T, et al.: Brain from bone: Efficient “meta-differentiation” of marrow stroma-derived mature osteoblasts to neurons with Noggin or a demethylating agent. Differentiation. 2001; 68: 235-244.

Song S, Song S, Zhang H, et al.: Comparison of neuron-like cells derived from bone marrow stem cells to those differentiated from adult brain neural stem cells. Stem Cells Dev. 2007; 16: 747-756.

Liu XP, Zhang ZQ, Gao SZ, et al.: Experimental study on the effect of mobilizing autologous bone marrow stem cells on focal cerebral ischemia/reperfusion injury and neuron apoptosis in rat. Chin J Neurosci. 2004; 20: 226-230.

Wells WA: Is transdifferentiation in trouble? J Cell Biol. 2002; 157: 15-18.

Li XZ, Bai LM, Yang YP, et al.: Effects of IL-6 secreted from astrocytes on the survival of dopaminergic neurons in lipopolysaccharide-induced inflammation. Neurosci Res. 2009; 65: 252-258.

Bath PM, Sprigg N: Colony stimulating factors (including erythropoietin, granulocyte colony stimulating factor and analogues) for stroke. Cochrane Database Syst Rev. 2008: CD005207.

Venkataramana NK, Kumar SKV, Balaraju S, et al.: Open labeled study of unilateral autologous bone-marrow-derived mesenchymal stem cell transplantation in Parkinson’s disease. Transl Res. 2011; 155: 62-70.



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