A New Method of Isolating Spinal Motor Neurons from Fetal Mouse

Publication date: Available online 23 June 2017
Source:Journal of Neuroscience Methods
Author(s): Weifang Wang, Bao Qi, Hui Lv, Fei Wu, Lulu Liu, Wei Wang, Quanquan Wang, Liangchen Hu, Yanlei Hao, Yuzhong Wang
BackgroundIsolating of primary motor neurons from animal embryos is critical for the study of neurological disease including mechanistic discovery and therapeutic development. Density gradient centrifuge taking advantage of the buoyant of motor neuron permits the enrichment of motor neurons. Despite the metrizamide, an OptiPrep medium has been introduced to separate the motor neurons by gradient centrifuge.New methodWe hereby used single density gradient of OptiPrep medium to isolate the spinal motor neurons from the fetal mouse.ResultsSingle density gradient of OptiPrep medium is effective to isolate spinal motor neurons from the fetal mouse. The immunofluorescence staining analysis showed that the purity of cultured motor neurons at 72h was between 90% and 95%.Comparison with Existing MethodFour gradients of OptiPrep medium have been previously used to isolate the motor neurons from spinal cord of mouse. In this study, the single gradient of OptiPrep medium was demonstrated to effectively isolate spinal motor neurons from the fetal mouse.ConclusionsThe single gradient of OptiPrep medium is enough to produce high purity of spinal motor neurons from the fetal mouse.



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