Acta Neurobiol Exp 2013, 73: 102–115

Treating spinal cord injury in rats with a combination of human fetal neural stem cells and hydrogels modified with serotonin

Ruzicka J.,2, Romanyuk N.1, Hejcl A.1,3, Vetrik M.4,5, Hruby M.4, Cocks G.6, Cihlar J.7, Pradny M.4, Price J.6, Sykova E.1,2, and Jendelova P.1,2

1Institute of Experimental Medicine, ASCR, Prague, Czech Republic;
2Department ofNeuroscience, Charles University, 2nd Faculty of Medicine, Prague, Czech Republic;
3Department of Neurosurgery, Masaryk Hospital, Usti nad Labem, Czech Republic;  
4Institute of Macromolecular Chemistry ASCR, Prague, Czech Republic;
5Charles University, Faculty of Science, Prague, Czech Republic; 
6Institute of Psychiatry,Kings College London, United Kingdom;                                                                     
7Department of Mathematics, Faculty of Science, J.E. Purkynje University in Ústi nad Labem, Czech Republic



Currently, there is no effective strategy for the treatment of spinal cord injury (SCI). A combination of biomaterials and stem cell therapy seems to be a promising approach to increase regenerative potential after SCI. We evaluated the use of a cell-polymer construct based on a combination of the conditionally immortalized spinal progenitor cell line SPC-01_GFP3, derived from human fetal spinal cord tissue, with a serotonin-modified poly(2-hydroxyethyl methacrylate) hydrogel (pHEMA-5HT). We compared the effect of treatment with a pHEMA-5HT hydrogel seeded with SPC-01_GFP3 cells, treatment with a pHEMA-5HT only and no treatment on functional outcome and tissue reconstruction in hemisected rats. Prior to transplantation the cell-polymer construct displayed a high potential to support the growth, proliferation and differentiation of SPC-01 cells in vitro. One month after surgery, combined hydrogel-cell treatment reduced astrogliosis and tissue atrophy and increased axonal and blood vessel ingrowth into the implant; however, two months later only the ingrowth of blood vessels remained increased. SPC-01_GFP3 cells survived well in vivo and expressed advanced markers of neuronal differentiation. However, a majority of the transplanted cells migrated out of the lesion and only rarely remained in the hydrogel. No differences among the groups in motor or sensory recovery were observed. Despite the support of the hydrogel as a cell carrier in vitro, and good results in vivo one month postsurgery, there was only a small effect on long term recovery, mainly due to the limited ability of the hydrogels to support the in vivo growth and differentiation of cells within the implant. Further modifications will be necessary to achieve stable long term improvement in functional outcome.

PMID: 23595287



Figure shows the influence of implanted pHEMA hydrogel with serotonin molecule modified surface on SPC-01 human fetal neural precursor cells differentiation. One month after implantation the SPC-01 cells seeded in our hydrogel material showed in some aspects more mature phenotype then three months after implantation when applied
only alone without modified hydrogel. Seeded cells survived well, mostly at the hydrogel-tissue border and in the peripheral part of the implanted hydrogel. The implanted cells were positive for the neuronal markers nestin (A) and βIII-tubulin (B) as well as for glial marker GFAP (C) and neuronal mature markers ChAT (D) and Tau (E). The implanted cells also showed an astrocyte- like morphology (F). Scale bar is 50 μm (C, E), 20 μm (A, B), and 10 μm (D, F). Our hydrogel modification may serve as important modulatory factor in further human neural stem cell application. This modification could shorten their naturally long time necessary for differentiation and get earlier mature neurons and glial cells in vivo. SPC-01 neural stem cells were proved as a stable source of stem cells, ideal for their abilities to survive, proliferate, and differentiate in vitro and in vivo in different models of spinal cord injury.

Jiri Ruzicka-1

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