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Adipose-Derived Stem Cells Expressing the Neurogenin-2 Promote Functional Recovery After Spinal Cord Injury in Rat

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Abstract

Neurogenin2 (Ngn2) is a proneural gene that directs neuronal differentiation of progenitor cells during development. This study aimed to investigate whether the use of adipose-derived stem cells (ADSCs) over-expressing the Ngn2 transgene (Ngn2–ADSCs) could display the characteristics of neurogenic cells and improve functional recovery in an experimental rat model of SCI. ADSCs from rats were cultured and purified in vitro, followed by genetically modified with the Ngn2 gene. Forty-eight adult female Sprague–Dawley rats were randomly assigned to three groups: the control, ADSCs, and Ngn2–ADSCs groups. The hind-limb motor function of all rats was recorded using the Basso, Beattie, and Bresnahan locomotor rating scale for 8 weeks. Moreover, hematoxylineosin staining and immunohistochemistry were also performed. After neural induction, positive expression rate of NeuN in Ngn2–ADSCs group was upon 90 %. Following transplantation, a great number of ADSCs was found around the center of the injury spinal cord at 1 and 4 weeks, which improved retention of tissue at the lesion site. Ngn2–ADSCs differentiated into neurons, indicated by the expression of neuronal markers, NeuN and Tuj1. Additionally, transplantation of Ngn2–ADSCs upregulated the trophic factors (brain-derived neurotrophic factor and vascular endothelial growth factor), and inhibited the glial scar formation, which was indicated by immunohistochemistry with glial fibrillary acidic protein. Finally, Ngn2–ADSCs-treated animals showed the highest functional recovery among the three groups. These findings suggest that transplantation of Ngn2-overexpressed ADSCs promote the functional recovery from SCI, and improve the local microenvironment of injured cord in a more efficient way than that with ADSCs alone.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No. 81171147), “Xingwei Project” Key Personal Medical Research Foundation of Health Department of Jiangsu Province (No. RC201156), “Six Categories of Key Person” Research Foundation of Jiangsu Province (No. 069), Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. JX10231801), and the science and technology plan projects of Jintan (JT2014059).

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    1. Department of Neurosurgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, Jiangsu, China

      Linjun Tang, Ronglan Zhu, Yi Tao, Kai Li, Jinyu Zheng, Penglai Zhao, Shuai Li, Xi Wang & Lixin Li

    2. Department of  Neurosurgery, Tongling Municipal Hospital, Tongling, 244000, Anhui, China

      Linjun Tang

    3. Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China

      Xiaocheng Lu

    4. Department of Neurosurgery, Jiangsu University Affiliated Jintan Hospital, Jintan, 213200, Jiangsu, China

      Tengda Qian

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    Linjun Tang, Xiaocheng Lu, and Ronglan Zhu have contributed equally to this work.

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    Supplementary Fig. 1

    Immunocytochemistry to detect the expressions of neural marker in GFP-ADSCs, expression of NeuN (A) and Tuj1 (B) in control medium or NeuN (C) and Tuj1 (D) neural induction medium for 7 days. Scale bars represent 100 um. Supplementary material 1 (TIFF 163 kb)

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    Tang, L., Lu, X., Zhu, R. et al. Adipose-Derived Stem Cells Expressing the Neurogenin-2 Promote Functional Recovery After Spinal Cord Injury in Rat. Cell Mol Neurobiol 36, 657–667 (2016). https://doi.org/10.1007/s10571-015-0246-y

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