24-well Plate Preparation for High and Low Density Neuron Culture
1:48
Brain Removal and Hippocampi Dissection of E16.5 - E17.5 Mouse Embryos
4:02
Enzymatic Digestion and Separation into Single Neurons
6:53
Plating of Neurons and Long Term Co-Culture
8:51
Results: Experimental Manipulations of Low Density Neurons Grown in Co-Culture
10:23
Conclusion
TRANSCRIÇÃO
The overall goal of this procedure is to establish healthy longterm cultures of primary mouse hippocampal neurons at ultra-low density, in order to facilitate immunocytochemistry labeling, and the study of neuronal cell autonomous mechanisms. This
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Low density cultures of primary hippocampal neurons usually require glia feeder layer to supply neurotrophic factors and sustain longevity. We describe here a simplified method to culture ultra-low density neurons on glass coverslips in the presence of a high density neuronal feeder layer, which facilitates investigation of specific neuronal-autonomous mechanisms.