Transdifferentiation of canine mesenchymal stem cells into neuron-like cells by induction with Î²-mercaptoethanol

BHABESH MILI; KINSUK DAS; MADHUSOODAN A P; KULDEEP KUMAR; A C SAXENA; SADHAN BAG

doi:10.56093/ijans.v91i7.115900

Authors

BHABESH MILI ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
KINSUK DAS ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
MADHUSOODAN A P ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
KULDEEP KUMAR ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
A C SAXENA ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
SADHAN BAG ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India

https://doi.org/10.56093/ijans.v91i7.115900

Keywords:

Î²-mercaptoethanol (BME), Canine, Mesenchymal stem cells, Neuron-like cells

Abstract

The objective of this study was to check whether β- mercaptoethanol in a culture medium can induce the neuronal differentiation of canine MSCs. The canine bonemarrow derived MSCs were first pre-inducted with 1 mM BME for 24 hrs followed by induction in a serum-free medium supplemented with 4 mM BME without FBS for another 6 days. Morphological changes in MSCs from spindle-shaped to neuron-like branching from the edges of the cells were noticed at the end of induction. These neuronlike cells were found positive for the immunophenotypic expression of different neural cell markers β-tubulin III, MAP-2 and Nestin. In RT-PCR analysis, it was also evident that the relative expressions of these representative genes were significantly higher in the differentiated cells. On the basis of our observations, it can be summarized that the BME induction of canine MSCs resulted in morphological changes that resembled neuron-like cells which were found to express the representative neuronal markers. Therefore, inducing canine MSCs with BME resulted in the generation of neuron-like cells that might be utilized for the prospective therapeutic applications in veterinary medicine.

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References

Bhat I A, Sivanarayanan T, Somal A, Pandey S, Bharti M K, Panda B S K, Indu B, Verma M, Anand J, Sonwane A, Kumar G S, Amarpal and Sharma G T. 2018. An allogenic therapeutic strategy for canine spinal cord injury using mesenchymal stem cells. Journal of Cellular Physiology 1–14. DOI: https://doi.org/10.1002/jcp.27086

Chung C S, Fujita N, Kawahara N, Yui S, Nam E and Nishimura R. 2013. A comparison of neurosphere differentiation potential of canine bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells. Journal of Veterinary Medical Sciences75: 879–86. DOI: https://doi.org/10.1292/jvms.12-0470

Das K, Madhusoodan A P, Mili B, Kumar A, Saxena A C, Kumar K, Sarkar M, Singh P, Srivastaba S and Bag S. 2017. Functionalized carbon nanotubes as suitable scaffold materials for proliferation and differentiation of canine mesenchymal stem cells. International Journal of Nanomedicine 12: 3235–52. DOI: https://doi.org/10.2147/IJN.S122945

Deng J, Petersen B E, Steindler D A, Jorgensen M L and Laywell E D. 2006. Mesenchymal stem cells spontaneously express neural proteins in culture and are neurogenic after transplantation. Stem Cells 24(4): 1054–64. DOI: https://doi.org/10.1634/stemcells.2005-0370

Edamura K, Kuriyama K, Kato K, Nakano R, Teshima K, Asano K, Sato T and Tanaka S. 2012. Proliferation capacity, neuronal differentiation potency and microstructures after the differentiation of canine bone marrow stromal cells into neurons. Journal of Veterinary Medical Science 74: 923–27. DOI: https://doi.org/10.1292/jvms.11-0388

Hayase M, Kitada M, Wakao S, Itokazu Y, Nozaki K, Hashimoto N, Takaqi Y and Dezawa M. 2009. Committed neural progenitor cells derived from genetically modified bone marrow stromal cells ameliorate deficits in a rat model of stroke. Journal of Cerebral Blood Flow Metaolism 29(8):1409–20. DOI: https://doi.org/10.1038/jcbfm.2009.62

Heo J S, Choi S M, Kim HO, Kim H, You J, Park C T, Kim C E and Kim H S. 2013. Neural transdifferentiation of human bone marrow mesenchymal stem cells on hydrophobic polymermodified surface and therapeutic effects in an animal model of ischemic stroke. Neuroscience 238: 305–18. DOI: https://doi.org/10.1016/j.neuroscience.2013.02.011

Jiang J, Lv,Z, Gu Y, Li J, Xu L, Xu W, Lu J, Xu J. 2010. Adult rat mesenchymal stem cells differentiate into neuronal-like phenotype and express a variety of neuro-regulatory molecules in vitro. Neuroscience Research 66: 46–52. DOI: https://doi.org/10.1016/j.neures.2009.09.1711

Jung D, Ha J, Kang B T, Kim J W, Quan F S, Lee J H, Woo E J and Park H M. 2009. A comparison of autologous and allogenic bone marrow derived mesenchymal stem cell transplantation in caninespinal cord injury. Journal of Neuronal Sciences 285: 67–77. DOI: https://doi.org/10.1016/j.jns.2009.05.027

Kim E Y, Lee K B, Yu J, Lee J H, Kim K J, Han K W, Park K S, Lee D and Kim M K. 2014. Neuronal cell differentiation of mesenchymal stem cells originating from canine amniotic fluid. Human Cell 27: 51–58. DOI: https://doi.org/10.1007/s13577-013-0080-9

Mili B, Das K, Kumar A, Saxena A C, Singh P, Ghosh S and Bag S. 2018. Preparation of NGF encapsulated chitosan nanoparticles and its evaluation on neuronal differentiation potentiality of canine mesenchymal stem cells. Journal of Material Science: Material Medicine 29: 4. DOI: https://doi.org/10.1007/s10856-017-6008-2

Pfaffl M W. 2001. A new mathematical model for relative quantification in real time-PCR. Nucleic Acid Research 29: 2002–07. DOI: https://doi.org/10.1093/nar/29.9.e45

Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman T B, Saporta S, Janssen W, Patel N, Cooper D R and Sanberg P R. 2000. Adult bone marrow stromal cells differentiate into neural cells in vitro. Experimental Neurology 164: 247–56. DOI: https://doi.org/10.1006/exnr.2000.7389

Vieira N M, Brandalise V, Zucconi E, Secco M B, Strauss B E and Zatz M. 2010. Isolation, characterization and differentiation potential of canine adipose-derived stem cell. Cell Transplantation 19: 279–89. DOI: https://doi.org/10.3727/096368909X481764

Wilcox J T, Lai J K Y, Semple E, Brisson B A, Gartley C, Armstrong J N, Betts D H 2011. Synaptically-competent neurons derived from canine embryonic stem cells by lineage selection with EGF and Noggin. PLoS One 6: e19786. DOI: https://doi.org/10.1371/journal.pone.0019768

William J B, Prabakaran R, Ayyappan S, Puskhinraj H, Rao D, Rao S, Manjunath Thamaraikannan P, Dedeepiya V D, Kuroda S, Yoshioka H, Mori Y, Preethy S and Abraham S J K. 2011. Functional recovery of spinal cord injury following application of intralesional bone marrow mononuclear cells embedded in polymer scaffold – two year follow-up in a canine. Journal of DOI: https://doi.org/10.4172/2157-7633.1000110

Stem Cell Research and Therapy 1: 3.

Woodbury D, Schwarz E J, Prockop D J and Black I B. 2000. Adult rat and human bone marrow stromal cells differentiate into neurons. Journal of Neuroscience Research 61: 364–70. DOI: https://doi.org/10.1002/1097-4547(20000815)61:4<364::AID-JNR2>3.0.CO;2-C

Wright K T, Masri W, Osman A, Chowdhury J. and Johnson W E. 2012. Concise review: Bone marrow for the treatment of spinal cord injury: mechanisms and clinical applications. Stem Cells 29: 169–78. DOI: https://doi.org/10.1002/stem.570

Yang Q, Mu J, Li Q, Li A, Zeng Z, Yang J, Zhang X, Tang J. and Xie P 2008. A simple and efficient method for deriving neurospheres from bone marrow stromal cells. Biochemical Biophysis Research Communication 372: 520–24. DOI: https://doi.org/10.1016/j.bbrc.2008.05.039