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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 10 (october 2019) : 1335-1339,   Doi: 10.18805/ijar.B-3647

Isolation and characterization of neural stem cells from ovine foetal cerebral cortex

S
S. Lija
S
S. Eswari
M
M. Monisha
K
K. Vijayarani
1<div style="text-align: justify;">Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 007, Tamil Nadu, India.</div>
Cite article:- Lija S., Eswari S., Monisha M., Vijayarani K. (2018). Isolation and characterization of neural stem cells from ovine foetal cerebral cortex. Indian Journal of Animal Research. 53(10): 1335-1339. doi: 10.18805/ijar.B-3647.

ABSTRACT

Neural stem cells are a potential source in cell based therapy for neurodegenerative diseases and traumatic brain injuries. The aim of this study is to isolate the ovine (Ovis aries) fetal neural stem cells and to analyze their neurogenic characteristics in vitro. Cerebral cortex pieces harvested from ovine fetal brain tissue were triturated for dispersal into single cells, cultured in neuronal stem cells culture medium and expanded in vitro in adherent culture. The morphological changes and the expression of neuron-specific gene markers and proteins were evaluated. These isolated NSCs were plastic adherent, self-renewing cells maintained the capacity to differentiate into brain specific cell types and they were found positive for neuronal stem cell markers, Nestin and â III tubulin by Reverse Transcription-PCR. Immunocytochemistry of the isolated NSCs also showed positive for Nestin, an established marker for neural and glial precursors as well as MSC marker CD90 and negative for  hematopoietic CD34 markers. 

REFERENCES

  1. Agarwal, P., Manish, K., Kuldeep, K., Renu, S., Puspendra, S. M., Ajay, K., Sanjeev, K. B., Dhruba, M., Mihir, S and Bag, S. (2014). Isolation and propagation of neural stem cells in caprine (Capra hircus).Cell Biology International, doi:10.1002/cbin.10282.
  2. Behnan,. J, Biljana, S, Tiziana, L., Gaetano, F., Giovanni, T., Torstein, R. M and Wayne, M. (2017). Identification and characterization of a new source of adult human neural progenitors. Cell Death and Disease, doi:10.1038/cddis.2017.368.
  3. Fu, Y.S., Cheng, Y. C., Lin, M.A., Cheng, H., Chu, P., Chou, S., Shih, Y., Kom and Sung, M. (2006). Conversion of human umbilical cord mesenchymal stem cells in wharton’s jelly to dopaminergic neurons in vitro: Potential therapeutic application for parkinsonism. Stem Cells, 24:115–124.
  4. Gade, N.E., Amar Nath, Pratheesh, M.D., Dubey, P.K., Amarpal, Sai Kumar G and Taru Sharma, G, (2012). Stem cell therapy in animal sciences – A review. Agri. Review, 33 (2) : 150 – 158.
  5. Kumar, K., Singh, R., Kumar, M., Agarwal, P., Mahapatra, P.S., Kumar, A., Malakar, D and Bag, S. (2013). Isolation and characterization of neural stem cells from buffalo. Journal of Neuroscience, doi: 10.3109/00207454.2013.852087
  6. Li, Q., Zhang, S., Zheng, S., Wen, H., Han, X., Zhang, M and Guan, W. (2017). Differentiation potential of neural stem cells derived from fetal sheep. Animal Cells and Systems, 21:4, 233-240.
  7. Li, .X, Xu, J and Bai, Y. (2005). Isolation and characterization of neural stem cells from human fetal striatum. Biochemistry and Biophysics Research Communication., 326:S425–434.
  8. Lindvall, O., Barker, R.A., Brustle, O., Isacson, O and Svendsen, C.N. (2012). Clinical translation of stem cells in neurodegenerative disorders. Cell Stem Cell.,10:151–155.
  9. Liu, Z., Wang, W., Gao, J., Zhou, H and Zhang, Y. (2014). Isolation, culture, and induced multiple differentiation of Mongolian sheep bone marrow-derived mesenchymal stem cells.In Vitro Cellular & Developmental Biology – Animal, 50(5): 464–474 
  10. Marei, H.E., Ahmed, A.E., Michetti, F., Pescatori, M., Pallini, R., Casalbore, P., Cenciarelli, C and Elhadidy, M. (2012). Gene expression profile of adult human olfactory bulb and embryonic neural stem cell suggests distinct signalling pathways and epigenetic control. PLoS One, 7:e33542.
  11. Murrell, W., Palmero, E., Bianco, J., Stangeland, B., Joel, M and Paulson, L. (2013). Expansion of multipotent stem cells from the adult human brain. PLoS One ,8: e71334.
  12. Osterberg, N and Roussa, E. (2009). Characterization of primary neurospheres generated from mouse ventral rostral hindbrain. Cell Tissue Research, 336:11–20.
  13. Zhao, M.T., Yang, X., Lee, K., Mao, J., Teson, J.M., Whitworth, K.M., Samuel, M.S., Spate, L.D., Murphy, C.N., Prather, R.S. (2012). The in vivo developmental potential of porcine skin-derived progenitors and neural stem cells. Stem Cells Dev 21: 2682–    2688.
  14. Zheng, Y.M., Dang, Y.H., Qiu, S., Qi, Y.P., Xu, Y.P., Sai, W.J. (2011). Differentiation and characteristics of the enhanced green fluorescent protein gene transgenic goat neural stem cells cultured in attached and non-attached plates. Cell Biol Int . 35: 849–856. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 10 (october 2019) : 1335-1339,   Doi: 10.18805/ijar.B-3647

    Isolation and characterization of neural stem cells from ovine foetal cerebral cortex

    S
    S. Lija
    S
    S. Eswari
    M
    M. Monisha
    K
    K. Vijayarani
    1<div style="text-align: justify;">Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 007, Tamil Nadu, India.</div>
    Cite article:- Lija S., Eswari S., Monisha M., Vijayarani K. (2018). Isolation and characterization of neural stem cells from ovine foetal cerebral cortex. Indian Journal of Animal Research. 53(10): 1335-1339. doi: 10.18805/ijar.B-3647.

    ABSTRACT

    Neural stem cells are a potential source in cell based therapy for neurodegenerative diseases and traumatic brain injuries. The aim of this study is to isolate the ovine (Ovis aries) fetal neural stem cells and to analyze their neurogenic characteristics in vitro. Cerebral cortex pieces harvested from ovine fetal brain tissue were triturated for dispersal into single cells, cultured in neuronal stem cells culture medium and expanded in vitro in adherent culture. The morphological changes and the expression of neuron-specific gene markers and proteins were evaluated. These isolated NSCs were plastic adherent, self-renewing cells maintained the capacity to differentiate into brain specific cell types and they were found positive for neuronal stem cell markers, Nestin and â III tubulin by Reverse Transcription-PCR. Immunocytochemistry of the isolated NSCs also showed positive for Nestin, an established marker for neural and glial precursors as well as MSC marker CD90 and negative for  hematopoietic CD34 markers. 

    REFERENCES

    1. Agarwal, P., Manish, K., Kuldeep, K., Renu, S., Puspendra, S. M., Ajay, K., Sanjeev, K. B., Dhruba, M., Mihir, S and Bag, S. (2014). Isolation and propagation of neural stem cells in caprine (Capra hircus).Cell Biology International, doi:10.1002/cbin.10282.
    2. Behnan,. J, Biljana, S, Tiziana, L., Gaetano, F., Giovanni, T., Torstein, R. M and Wayne, M. (2017). Identification and characterization of a new source of adult human neural progenitors. Cell Death and Disease, doi:10.1038/cddis.2017.368.
    3. Fu, Y.S., Cheng, Y. C., Lin, M.A., Cheng, H., Chu, P., Chou, S., Shih, Y., Kom and Sung, M. (2006). Conversion of human umbilical cord mesenchymal stem cells in wharton’s jelly to dopaminergic neurons in vitro: Potential therapeutic application for parkinsonism. Stem Cells, 24:115–124.
    4. Gade, N.E., Amar Nath, Pratheesh, M.D., Dubey, P.K., Amarpal, Sai Kumar G and Taru Sharma, G, (2012). Stem cell therapy in animal sciences – A review. Agri. Review, 33 (2) : 150 – 158.
    5. Kumar, K., Singh, R., Kumar, M., Agarwal, P., Mahapatra, P.S., Kumar, A., Malakar, D and Bag, S. (2013). Isolation and characterization of neural stem cells from buffalo. Journal of Neuroscience, doi: 10.3109/00207454.2013.852087
    6. Li, Q., Zhang, S., Zheng, S., Wen, H., Han, X., Zhang, M and Guan, W. (2017). Differentiation potential of neural stem cells derived from fetal sheep. Animal Cells and Systems, 21:4, 233-240.
    7. Li, .X, Xu, J and Bai, Y. (2005). Isolation and characterization of neural stem cells from human fetal striatum. Biochemistry and Biophysics Research Communication., 326:S425–434.
    8. Lindvall, O., Barker, R.A., Brustle, O., Isacson, O and Svendsen, C.N. (2012). Clinical translation of stem cells in neurodegenerative disorders. Cell Stem Cell.,10:151–155.
    9. Liu, Z., Wang, W., Gao, J., Zhou, H and Zhang, Y. (2014). Isolation, culture, and induced multiple differentiation of Mongolian sheep bone marrow-derived mesenchymal stem cells.In Vitro Cellular & Developmental Biology – Animal, 50(5): 464–474 
    10. Marei, H.E., Ahmed, A.E., Michetti, F., Pescatori, M., Pallini, R., Casalbore, P., Cenciarelli, C and Elhadidy, M. (2012). Gene expression profile of adult human olfactory bulb and embryonic neural stem cell suggests distinct signalling pathways and epigenetic control. PLoS One, 7:e33542.
    11. Murrell, W., Palmero, E., Bianco, J., Stangeland, B., Joel, M and Paulson, L. (2013). Expansion of multipotent stem cells from the adult human brain. PLoS One ,8: e71334.
    12. Osterberg, N and Roussa, E. (2009). Characterization of primary neurospheres generated from mouse ventral rostral hindbrain. Cell Tissue Research, 336:11–20.
    13. Zhao, M.T., Yang, X., Lee, K., Mao, J., Teson, J.M., Whitworth, K.M., Samuel, M.S., Spate, L.D., Murphy, C.N., Prather, R.S. (2012). The in vivo developmental potential of porcine skin-derived progenitors and neural stem cells. Stem Cells Dev 21: 2682–    2688.
    14. Zheng, Y.M., Dang, Y.H., Qiu, S., Qi, Y.P., Xu, Y.P., Sai, W.J. (2011). Differentiation and characteristics of the enhanced green fluorescent protein gene transgenic goat neural stem cells cultured in attached and non-attached plates. Cell Biol Int . 35: 849–856. 
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