Investigation of Neuronal Phenotype Injection in P19 Stem Cells Under the Influence of Rat Infant Extract and Deprenil

Document Type : Original Article

Authors

1 Department of Biology, Faculty of Sciences, University of Isfahan, Hezarjerib Avenue, Isfahan 8174673441, Iran

2 Depaartment of biology, faculty of sciences, University of Isfahan, Hezarjerib Avenue, Isfahan 8174673441, Iran

Abstract

Introduction: Parkinson's disease cause by selective loss of a group of dopaminergic cells in the Substantial nigra. One of the treatments for this disease is the displacement of lost cells by stem cells. The purpose of this study was to investigate the effect of inducers of extract of neonatal rat brain and deprenyl on neural differentiation of P19 embryonic carcinoma cells.
Materials and Methods: P19 stem cells were cultured in alpha-MEM medium supplemented with 10% FBS serum. Then, the cells were cultured on low-adhesion culture plates to create pseudo-embryonic bodies. In order to differentiate, germ cells were cultured in medium containing 3% FBS serum and extract of neonatal rat brain, deprenyl and mixture of both extract for 28 days. Immunofluorescence assay and Real-time PCR was applied to detect and evaluate the expression specific proteins of neurons including synaptophysin, beta-tubulin 3 and nestin, respectively.
Results: Regarding to immunofluorescence results, differentiated P19 cells, positively affected by three inducers such as nestin, synaptophysin and beta-3. Relative expression of transcription factors of nestin, synaptophysin in cells differentiated by all three inducer using Real-time PCR demonstrated that in P19 embryonic carcinoma cells, Nestin and Synaptophysin expression increased in the second week, while, thier expression significantly reduced in the third and fourth weeks.
Conclusion: The results of this study indicated P19 embryonic carcinoma stem cells are potential to differentiate into neurons, especially dopaminergic phenotype, following laboratory conditions and inducing factors

Keywords

References

1-D'Amour K, Gage FH. New tools for human developmental biology. Nature biotechnology. 2000;18(4):381.
2-Du Y, Funderburgh ML, Mann MM, SundarRaj N, Funderburgh JL. Multipotent stem cells in human corneal stroma. Stem cells. 2005;23(9):1266-75.
3-Tuch BE. Stem cells: a clinical update. Australian family physician. 2006;35(9):719.
4-Dantuma E, Merchant S, Sugaya K. Stem cells for the treatment of neurodegenerative diseases. Stem cell research & therapy. 2010;1(5):37.
5-Hsieh FF, Barnett LA, Green WF, Freedman K, Matushansky I, Skoultchi AI, et al. Cell cycle exit during terminal erythroid differentiation is associated with accumulation of p27Kip1 and inactivation of cdk2 kinase. Blood. 2000;96(8):2746-54.
6-Choi S-C, Choi J-H, Shim W-J, Lim D-S. P19 embryonal carcinoma cells: a new model for the study of endothelial cell differentiation. Biotechnology letters. 2008;30(7):1169-75.
7-Iwasaki Y, Ikeda K, Shiojima T, Kobayashi T, Tagaya N, Kinoshita M. Deprenyl enhances neurite outgrowth in cultured rat spinal ventral horn neurons. Journal of the neurological sciences. 1994;125(1):11-3.
8-Sweetman SC. Martindale: the complete drug reference2005.
9-Brannan T, Yahr MD. Comparative study of selegiline plus l‐dopa–carbidopa versus l‐dopa–carbidopa alone in the treatment of parkinson's disease. Annals of Neurology: Official Journal of the American Neurological Association and the Child Neurology Society. 1995;37(1):95-8.
10-Tatton W, Wadia J, Ju W, Chalmers-Redman R, Tatton N. (−)-Deprenyl reduces neuronal apoptosis and facilitates neuronal outgrowth by altering protein synthesis without inhibiting monoamine oxidase.  Deprenyl—Past and Future: Springer; 1996. p. 45-59.
11-Mayanil C, Baquer N. Comparison of the properties of semipurified mitochondrial and cytosolic monoamine oxidases from rat brain. Journal of neurochemistry. 1984;43(4):906-12.
12-Yoshikawa K, Aizawa T, Hayashi Y. Degeneration in vitro of post-mitotic neurons overexpressing the Alzheimer amyloid protein precursor. Nature. 1992;359(6390):64.
13-Bakhshalizadeh S, Esmaeili F, Houshmand F, Shirzad H, Saedi M. Effects of selegiline, a monoamine oxidase B inhibitor, on differentiation of P19 embryonal carcinoma stem cells, into neuron-like cells. In Vitro Cellular & Developmental Biology-Animal. 2011;47(8):550.
14-McBurney MW, Jones-Villeneuve E, Edwards M, Anderson P. Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature. 1982;299(5879):165.
15-Esmaeili F, Tiraihi T, Movahedin M, Mowla SJ. Selegiline induces neuronal phenotype and neurotrophins expression in embryonic stem cells. Rejuvenation research. 2006;9(4):475-84.
16-Lendahl U, Zimmerman LB, McKay RD. CNS stem cells express a new class of intermediate filament protein. Cell. 1990;60(4):585-95.
17-Falconer MM, Echeverri CJ, Brown DL. Differential sorting of beta tubulin isotypes into colchicine‐stable microtubules during neuronal and muscle differentiation of embryonal carcinoma cells. Cell motility and the cytoskeleton. 1992;21(4):313-25.
18-McBurney MW, Reuhl K, Ally A, Nasipuri S, Bell J, Craig J. Differentiation and maturation of embryonal carcinoma-derived neurons in cell culture. The Journal of neuroscience. 1988;8(3):1063-73.
19-Wiese C, Rolletschek, A., Kania, G., Blyszczuk, P., Tarasov, K. V., Tara-sova, Y., Wersto, R. P., Boheler, K. R., and Wobus, A. M. Cell MolLife Sci. 2004:61, 2510–22.
20-Esmaeili F, Tiraihi, T, Movahedin, M, Mowla SJ. Selegiline induces neuronal phenotype and neurotrophins expression in embryonic stem cells. Rejuvenation Res. 2006: 9(4): 475-84.
21-McBurney MW. Clonal lines of teratocarcinoma cells in vitro: differentiation and cytogenetic characteristics. J Cell Physiol. 1976:89(3): 441-55.
22-Vannucchi MG F-PM. Synapse formation during neuron differentiation:an in situ study of the myenteric plexus during murine embryonic life. J Comp Neurol. 2000:425(3): 369-81.
23-Sortwell CE ea. Pattern of synaptophysin immunoreactivity within mesencephalic grafts following transplantation in a parkinsonian primate model.    . Brain Res. 1998:791(1): 117-24.
Jundishapur Scientific Medical Journal
Volume 19, Issue 1 - Serial Number 124
March and April 2021
Pages 83-95

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History

  • Receive Date: 01 August 2019
  • Revise Date: 21 February 2020
  • Accept Date: 29 February 2020

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