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CJAP ›› 2017, Vol. 33 ›› Issue (5): 445-449.doi: 10.12047/j.cjap.5563.2017.107

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Establishment of neuritis migration model induced by Slit 2N and silk fibroin mixture in the rat hippocampal neurons in vitro

GANG Lin1,2, LI Yi-peng1,2, LU Lei1,2, YANG Kai2,3, SUN Zhong-lei2,3, CHEN Xu-yi2, TU Yue1,2   

  1. 1. Tianjin University of Traditional Chinese Medicine, Tianjin 300193;
    2. Tianjin Key Laboratory of Neurotrauma Repair, Institute of Traumatic Brain Injury and Neurology, Neurological Department of Affiliated Hospital of Logistics University of Chinese Armed Police Forces, Tianjin 300162;
    3. Jinzhou Medical University, Jinzhou 121001, China
  • Received:2017-02-04 Revised:2017-04-27 Online:2017-09-28 Published:2018-06-19
  • Supported by:
    国家自然科学基金青年项目(11102235);天津市科技支撑计划重点项目(14ZCZDGX00500);天津市卫生局科技基金项目(2013KZ134,2014KZ135)

Abstract: Objective: To explore a simply feasible and affordable method to establish neuritis migration model induced by Slit 2N and silk fibroin mixture in the rat hippocampal neurons in vitro.Methods: Neurons were derived from SD rat hippocampal tissues and cultured with a special cell culture-plate in vitro. The cultured neurons were divided into four groups, named as control group, pure silk fibroin, pure Slit 2N and Slit 2N mixture with silk fibroin (mixture group), and 50 different neurons were randomly selected in each group. Moreover, we photographed and recorded the soma coordinate and added the silk fibroin, Slit 2N and mixture to each neurite with a distance of 100 μm, except control group. Record again after 30 min. Property and positive rate of cells were identified by immunofluorescence staining.Results: Neurites of the pure Slit 2N group and the mixture group became shorter, and there was no significant change in the pure silk fibroin and control groups. The results showed that the average duration and length difference before and after changed were in descending order of mixture group, Slit 2N group, silk fibroin group (P<0.05), and the silk fibroin and control groups were no significant change (P>0.05). The positive rate of MAP-2 in four groups was more than 90%.Conclusion: There were no significant effects of Silk fibroin on induced by Slit 2N in rat hippocampal neuron migration. It had an effect on reducing Slit 2N diffusion rate and extending its working time. It provides an advantageous construction method in vitro on based 3D directed neural repair for the treatment of central nervous system diseases.

Key words: Slit 2N, silk fibroin, hippocampal neuron, neural guidance, neural migration, nerve repair, rat

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