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The isolation and cultivation of bone marrow stem cells and evaluation of differences for neural-like cells differentiation under the induction with neurotrophic factors

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Abstract

The bone marrow represents the most common source from which to isolate mesenchymal stem cells (MSCs). They can be obtained directly from patients and successfully induced to form various differentiated cell types. In addition, cell-based transplantation therapies have been proven to be promising strategies for curing disease of the nerve system. Therefore, it was particularly important to establish an easy and feasible method for the isolation, purification, and differentiation of bone marrow stromal cells (BMSCs). The aim of this study was to isolate and characterize putative bone marrow derived MSCs from Sprague–Dawley (SD) rats. Furthermore, differentiation effects were compared between the GDNF-induction group and the BDNF-induction group. Of these, BMSCs were isolated from the SD rats in a traditional manner, and identified based on plastic adherence, morphology, and surface phenotype assays. After induction with GDNF and BDNF, viability of BMSCs was detected by MTT assay and neuronal differentiation of BMSCs was confirmed by using immunofluorescence and Western blotting. Besides, the number of BMSCs that obviously exhibited neuronal morphology was counted and the results were compared between the GDNF-induction group and BDNF-induction groups. Our results indicate that direct adherence was a simple and convenient method for isolation and cultivation of BMSCs. Furthermore, BMSCs can be induced in vitro to differentiate into neuronal cells by using GDNF, which could achieve a more persistent and stable inducing effect than when using BDNF.

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Acknowledgments

This work was supported in part by a Grant of the National Natural Science Foundation of China (NSFC81071466).

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Authors and Affiliations

  1. Department of spine surgery, The clinical medicine college of Yangzhou University, Nantong west road no.98, Yangzhou, Jiangsu province, china

    Jian-Dong Yang, Jing-Cheng Wang & Xin-Min Feng

  2. The clinical medicine college of Yangzhou University, Huaihai road no.11, Yangzhou, Jiangsu province, China

    Cheng-Huang, Yi-Nan Li & Hai-Xiang Xiao

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  1. Jian-Dong Yang
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  2. Cheng-Huang
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  3. Jing-Cheng Wang
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  4. Xin-Min Feng
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  6. Hai-Xiang Xiao
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Correspondence to Cheng-Huang.

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Yang, JD., Cheng-Huang, Wang, JC. et al. The isolation and cultivation of bone marrow stem cells and evaluation of differences for neural-like cells differentiation under the induction with neurotrophic factors. Cytotechnology 66, 1007–1019 (2014). https://doi.org/10.1007/s10616-013-9654-3

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