探讨阿糖胞苷在原代大鼠海马神经元培养中的纯化方法

doi:10.13418/j.issn.1001-165x.2015.06.014

Chinese Journal Of Clinical Anatomy ›› 2015, Vol. 33 ›› Issue (6): 681-684.doi: 10.13418/j.issn.1001-165x.2015.06.014

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Application of Ara-C in rat hippocampal neuron purificationculture

LI Ang¹, YAN Fang¹, LI Zhen-lin¹, LIU Zhong-ying¹, ZHANG Lin¹, DONG Wei-ren¹, NIU Hong-xin², WANG Hai-Hong¹

1. Department of Histology and Embryology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; 2. Department of Nephrology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, China

Received:2015-08-21 Online:2015-11-25 Published:2015-12-18

Abstract

Abstract:

Objective　To investigate the optimum condition of Ara-C treatment in cell purifica-tionculture of rat primary hippocampal neurons. Methods Primary neurons in hippocampal region of new born (1 day after birth) SD rats were cultured for 48 h and then were treated with different doses (0 mol/L, 2.5 mol/L, 5.0 mol/L, 7.5 mol/L and 10 mol/L) of Ara-C for 0 h, 6 h, 12 h and 24 h. Cellular morphology was observed by microscope. Cell viabilities were determined by MTT assay and neuron purities were calculated by morphology. To further assess the neuron purities, MAP2 antibody were used by immunofluorescence and western blot and cell viability is measured by Trypan blue staining. Results Hippocampal neuron purity and cell viability was optimal when 5 mol/L Ara-C was added for 24 h. Neuron purity with Ara-C was better than control (74.28±10.13)% VS (34.82±8.15)%, (P=0.000)). MAP2 protein level in cells treated with Ara-C was higher than that in control cells (P=0.008). The cell viabilities in both conditions were similar ((93.2±3.82)% for Ara-C treatment VS (95.6±2.37)% for control treatment, (P=0.109)). Conclusion The optimum condition of Ara-C treatment in terms of neuron purity and cell viability was 5 mol/L for 24 h in cell culture of rat primary hippocampal neurons.

Key words: Ara-C, Hippocampal Neurons, Purificationculture, Viability

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