Study of the method of spinal cord neuron culture in Sprague–Dawley rats

Ibrain Pub Date : 2022-12-25 DOI:10.1002/ibra.12085
Yi-Fei Sun, Quan-Yuan Chang, Narima Eerqing, Chang-Yan Hu
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Abstract

This study aimed to explore the method of culture of spinal cord neurons (SPNs) in vitro and to provide prerequisites for studying the molecular mechanism and pharmacological mechanism of spinal cord injury and repair. The spinal cord tissues of neonatal Sprague–Dawley rats were taken and digested by trypsin, followed by cytarabine (Ara-C) to inhibit the proliferation of heterogeneous cells, differential velocity adhesion, and natural growth in neuron-specific medium. Then, the morphology of SPNs was observed. Ara-C treatment inhibited the growth of heterogeneous cells and the growth of spinal neurons. Using the differential velocity adhesion method, it was found that the adhesion time of heterogeneous cells and SPNs was not significantly different, and it could not separate neurons and heterogeneous cells well. A large number of mixed cells gathered and floated, and died on the 18th day. Compared with the 20th day, the cell viability of the 18th day was better (p < 0.001). The natural growth and culture of SPNs in Neurobasal-A medium can yield neurons of higher purity and SPNs from the 12th day to the 18th day can be selected for related in vitro cell experiments.

Abstract Image

Sprague-Dawley大鼠脊髓神经元培养方法的研究。
本研究旨在探索脊髓神经元的体外培养方法,为研究脊髓损伤和修复的分子机制和药理学机制提供前提。取新生Sprague-Dawley大鼠的脊髓组织,用胰蛋白酶消化,然后用阿糖胞苷(Ara-C)在神经元特异性培养基中抑制异质细胞的增殖、差速粘附和自然生长。然后,观察SPN的形态。Ara-C处理抑制了异种细胞的生长和脊髓神经元的生长。采用差速粘附法,发现异种细胞和SPN的粘附时间没有显著差异,不能很好地分离神经元和异种细胞。大量混合细胞聚集漂浮,于第18天死亡。与第20天相比,第18天的细胞活力更好(p
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