ArticleJournal of visualized experiments : JoVE2016
A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture.
Article in Journal of visualized experiments : JoVE, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
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- Ex vivo engineering of neural tissue structure and growth using sequential 2D and 3D solid scaffolds.In vitro cellular & developmental biology. Animal · 2026Article
- A skin-specific α-Synuclein seeding amplification assay for diagnosing Parkinson's disease.NPJ Parkinson's disease · 2024Article
- Mouse serum albumin induces neuronal apoptosis and tauopathies.Acta neuropathologica communications · 2024Article
- Review
- Novel adult cortical neuron processing and screening method illustrates sex- and age-dependent effects of pharmaceutical compounds.Scientific reports · 2022Article
- Article
- Methyltransferase-Like 3 Rescues the Amyloid-beta protein-Induced Reduction of Activity-Regulated Cytoskeleton Associated Protein ExpressionFrontiers in aging neuroscience · 2022Article
- Neurotransmitter-stimulated neuron-derived sEVs have opposite effects on amyloid β-induced neuronal damage.Journal of nanobiotechnology · 2021Article
- MiR-132 down-regulates high glucose-induced β-dystroglycan degradation through Matrix Metalloproteinases-9 up-regulation in primary neurons.Journal of cellular and molecular medicine · 2021Article
- Review
- Inhibition of microRNA-129-2-3p protects against refractory temporal lobe epilepsy by regulating GABRA1.Brain and behavior · 2021Article
- Modeling the mature CNS: A predictive screening platform for neurodegenerative disease drug discovery.Journal of neuroscience methods · 2021Article
- Aragonite-Polylysine: Neuro-Regenerative Scaffolds with Diverse Effects on Astrogliosis.Polymers · 2020Article
- Autaptic Cultures: Methods and Applications.Frontiers in synaptic neuroscience · 2020Article
- Neurexin-1α regulates neurite growth of rat hippocampal neurons.International journal of physiology, pathophysiology and pharmacology · 2019Article
- Trkb-IP3 Pathway Mediating Neuroprotection in Rat Hippocampal Neuronal Cell Culture Following Induction of Kainic Acid.The Malaysian journal of medical sciences : MJMS · 2018Article
- Low-Density Neuronal Cultures from Human Induced Pluripotent Stem Cells.Molecular neuropsychiatry · 2017Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic hippocampal neurons can often grow successfully on glass coverslips at high density under serum-free conditions, but low density cultures typically require a supply of trophic factors by co-culturing them with a glia feeder layer, preparation of which can be time-consuming and laborious. In addition, the presence of glia may confound interpretation of results and preclude studies on neuron-specific mechanisms. Here, a simplified method is presented for ultra-low density (~2,000 neurons/cm2), long-term (>3 months) primary hippocampal neuron culture that is under serum free conditions and without glia cell support. Low density neurons are grown on poly-D-lysine coated coverslips, and flipped on high density neurons grown in a 24-well plate. Instead of using paraffin dots to create a space between the two neuronal layers, the experimenters can simply etch the plastic bottom of the well, on which the high density neurons reside, to create a microspace conducive to low density neuron growth. The co-culture can be easily maintained for >3 months without significant loss of low density neurons, thus facilitating the morphological and physiological study of these neurons. To illustrate this successful culture condition, data are provided to show profuse synapse formation in low density cells after prolonged culture. This co-culture system also facilitates the survival of sparse individual neurons grown in islands of poly-D-lysine substrates and thus the formation of autaptic connections.
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