Cell and Organ Transplantology. 2016; 4(1):22-28.
Transplantation of cryopreserved rat fetal neural cells in suspension and in multicellular aggregates into rats with spinal cord injury
Sukach A. N., Lebedinsky A. S., Ochenashko O. V., Petrenko A. Yu.
Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine
Today cell transplantation is one of the promising approaches of spinal cord injuries treatment. The aim of the work was to study the effect of cryopreserved fetal neural cell transplantation in suspensions and cell aggregates for motor activity recovery of rats with experimental spinal cord injury.
Materials and methods. Cells were isolated from the brain tissue of rat fetuses 15-16 days of gestation. The formation of aggregates was performed during short-term cultivation at a concentration of 8·106 cells/mL in medium with 10 % adult rat serum. Cell transplantation was performed into the damaged area of spinal cord in aggregates or suspension. To fix transplanted cells in the damaged area we used alginate gel.
Results. Transplantation of cryopreserved fetal neural cells in alginate gel had the positive effect on dynamics of rats’ motor activity recovery. That was manifested in the extensive mobility of three joints of one limb and the limited mobility of two joints of the other with simultaneous recovery of the sensitivity of the hind limbs.
Conclusion. Cryopreserved fetal neural cells aggregates had a high therapeutic potential on rat traumatic spinal cord injury compared with cell suspension by improving the structure of forming nervous tissue and significantly increasing the rate of hind limb function recovery.
Keywords: neural cell culture, cryopreservation of cells, spinal cord injury, cell transplantationFull Text PDF
1. Sukach АN, Grischenko VI. Kletochnaya terapiya neyrodegenerativnykh bolezney: istochniki kletok i strategiya ikh primeneniya [Cell therapy of neurodegenerative diseases: cell sources and strategy for their application]. Uspekhi sovremennoy biologii – Biology Bulletin Reviews. 2007; 127(1): 25-33 [in Russian].
|2. Emmerta MY, Hitchcock RW, Hoerstrupa SP. Cell therapy, 3D culture systems and tissue engineering for cardiac regeneration. Advanced Drug Delivery Reviews. 2014; 69–70: 254–69.
|3. Stabenfeldt SE, Munglani G, Garcia AJ, et al. Biomimetic Microenvironment Modulates Neural Stem Cell Survival, Migration, and Differentiation. Tissue engineering: Part A. 2010; 16(12): 3747- 58.
|4. Xu CJ, Wang JL, Jin WL. The Neural Stem Cell Microenvironment: Focusing on Axon Guidance Molecules and Myelin-Associated Factors. Journal of Molecular Neuroscience. 2015; 56(4,3): 887-97.|
|7. Lyashenko TD, Shevchenko MV. Svoystva izolirovannykh kletok nervnoy tkani novorozhdennykh krys v kul’ture [Properties of the isolated cells from nervous tissue of newborn rat in the culture]. Biotechnologia Acta. 2013; 6(3): 63-8 [in Russian].|
|8. Petrenko AYu, Sukach АN. Isolation of intact mitochondria and hepatocytes using vibration. Analytical Biochem. 1991; 194(2): 326-29.
|9. Sukach AN, Shevchenko MV, Liashenko TD. Comparative study of influence on fetal bovine serum and serum of adult rat on cultivation of newborn rat neural cells. Biopolymers and Cell. 2014; 30(5): 394-400.
|10. Woerly S, Doan VD, Evans-Martin F, et al. Spinal cord reconstruction using NeuroGelTM implants and functional recovery after chronic injury. J. of Neurosciensce Res. 2001; 66: 1187-97.
|11. Basso DM, Beatie MS, Bresnahan JC. Sensitive and reliable locomotor rating scale for open field testing in rats. Journal of neurotrauma. 1995; 12(1): 1-21.
|12. Leea KY, Mooneya DJ. Alginate: Properties and biomedical applications. Progress in Polymer Science. 2012; 37(1): 106–26.
|13. Lim F, Sun AM. Microencapsulated islets as bioartificial endocrine pancreas. F. Lim Science. 1980; 210(4472): 908-10.
|14. Goren A, Dahan N, Goren E, et al. Encapsulated human mesenchymal stem cells: a unique hypoimmunogenic platform for long-term cellular therapy. FASEB J. 2010; 24(1): 22-31.
|15. Gimi B, Nemani KV. Advances in alginate gel microencapsulation of therapeutic cells. Crit Rev Biomed Eng. 2013; 41(6): 469-81.
|16. Matyash M, Despang F, Mandal R, et al. Novel soft alginate hydrogel strongly supports neurite growth and protects neurons against oxidative stress. Tissue Engineering. Part A. 2012; 18(1-2): 55-66.
|17. Suzuki K, Suzuki Y, Ohnishi K, et al. Regeneration of transected spinal cord in young adult rats using freeze-dried alginate gel. NeuroReport. 1999; 10(14): 2891–94.
|18. Prang P, Muller R, Eljaouhari A, et al. The promotion of oriented axonal regrowth in the injured spinal cord by alginate-based anisotropic capillary hydrogels. Biomaterials. 2006; 27(19): 3560–69.
|19. Wu S, Suzuki Y, Kitada M, et al. Migration, integration, and differentiation of hippocampus-derived neurosphere cells after transplantation into injured rat spinal cord. Neuroscience Letters. 2001; 312(3): 173–76.
|20. Nori S, Okada Y, Yasuda A, et al. Grafted human-induced pluripotent stem-cell-derived neurospheres promote motor functional recovery after spinal cord injury in mice. Proc Natl Acad Sci USA. 2011; 108(40): 16825–830.
|21. Llado J, Haenggeli C, Maragakis NJ, et al. Neural stem cells protect against glutamate-induced excitotoxicity and promote survival of injured motor neurons through the secretion of neurotrophic factors. J. Mol Cell Neurosci. 2004; 27(3): 322-31.
|22. Lu P, Jones LL, Snyder EY, et al. Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury. Exp Neurol. 2003; 81(2): 115-29.
|23. Cao Q, He Q, Wang Y, et al. Transplantation of ciliary neurotrophic factor-expressing adult oligodendrocyte precursor cells promotes remyelination and functional recovery after spinal cord injury. J Neurosci. 2010; 30(8): 2989–3001.
|24. Heine W, Conant K, Griffin JW, et al. Transplanted neural stem cells promote axonal regeneration through chronically denervated peripheral nerves. Exp Neurol. 2004; 189(2): 231-40.
|25. Liu H, Shubayev V. Matrix metalloproteinase-9 controls proliferation of NG2+ progenitor cells immediately after spinal cord injury. Exp Neurol. 2011; 231(2): 236–46.
|26. Li X, Liu T, Song K, et al. Effect of neural stem cells on apoptosis of PC12 cells induced by serum deprivation. Biotechnol prog. 2007; 23(4): 952-57.
Usenko OYu, Yakushev AV, Kostylyev MV, Onischenko VF. Effect of transplantation of cord blood total nucleated cells on the manifestation and prognosis of refractory congestive heart failure. Cell and Organ Transplantology. 2016; 4(1):10-13. doi: 10.22494/COT.V4I1.6