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Updated: Mar 10, 2026
Low-Density Primary Hippocampal Neuron Culture
Published on: April 18, 2017
Protocol for culturing low density pure rat hippocampal neurons supported by mature mixed neuron cultures
Qian Yang1, Yini Ke1, Jianhong Luo1
1Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310058, China.
Background:
primary hippocampal neuron cultures allow for subcellular morphological dissection, easy access to drug treatment and electrophysiology analysis of individual neurons, and is therefore an ideal model for the study of neuron physiology. While neuron and glia mixed cultures are relatively easy to prepare, pure neurons are particular hard to culture at low densities which are suitable for morphology studies. This may be due to a lack of neurotrophic factors such as brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT3) and Glial cell line-derived neurotrophic factor (GDNF).
New Method:
In this study we used a two step protocol in which neuron-glia mixed cultures were initially prepared for maturation to support the growth of young neurons plated at very low densities.
Comparison With Existing Methods:
Our protocol showed that neurotrophic support resulted in physiologically functional hippocampal neurons with larger cell body, increased neurite length and decreased branching and complexity compared to cultures prepared using a conventional method.
Conclusion:
Our protocol provides a novel way to culture highly uniformed hippocampal neurons for acquiring high quality, neuron based data.
