The Thermo Scientific Pierce Primary Neuron Isolation Kit provides isolation and culturing reagents for the optimal yield and viability of primary neurons form embryonic cortical and hippocampal tissue from mouse and rat.
The Primary Neuron Isolation Kit has been designed to obtain high viability and purity of primary embryonic neurons using validated reagents and an optimized procedure for enzymatic digestion and culturing. The kit contains tissue-specific dissociation enzymes, media formulated specifically for neuronal cell culture, and cell culture supplements that promote the isolation and growth of functional neurons. The validated protocol and reagents produce highly pure primary neurons for either short- or long-term culture, and the kit is designed for use by both experienced and non-experienced users.
- Optimized – procedure and reagents optimized for viability, yield, purity, and ease-of-use
- Yield – provides a 2-fold increase in yield compared to do-it-yourself methods
- Viability – greater than 95% of isolated cells remain viable in culture from Day 1 to Week 4
- Functional – cultured neurons contain processes indicative of complete differentiation and functionality
- Complete kit includes Neuronal Isolation Enzyme, Hanks' Balanced Salt Solution (HBSS), Neuronal Culture Medium, Neuronal Culture Media Supplement, Glutamine Supplement, and Neuronal Growth Supplement
- Fresh embryonic mouse or rat tissue sample, heat-inactivated fetal bovine serum (FBS), poly-D-lysine treated culture dishes
- Neuronal cell differentiation
- Immunohistochemistry (IHC)
- Functional and biochemical assays
- Biologically relevant system for preclinical drug discovery, neurotoxicity testing, and predictive disease modeling
The Pierce Primary Neuron Isolation Kit has been optimized to provide excellent cell yield and to sustain cells at greater than 95% viability. The neurons are appropriately polarized, develop extensive axonal and dendritic arbors, express neuronal and synaptic markers, and form numerous, functional synaptic connections with one another. They can be used as a model system for molecular and cellular biology studies of neuronal development and function, especially for visualizing the subcellular localization of endogenous or expressed proteins, imaging protein, neuronal polarity, and dendritic growth and synapse formation .
|Cell yield and viability after cell isolation using the Thermo Scientific Pierce Neuron Isolation Kit. Mouse embryonic cortical tissue at E16-19 were incubated with Pierce neuronal isolation enzyme (with papain) for 30 minutes and disrupted by pipetting up and down with a pipette and 1000µL tip for 20 times to generate single cell suspension. Cell yield and cell viability are acquired from cell suspension isolated from one pair of mouse cortex in total volume of 1.5mL. Cell viability was determined by trypan blue exclusion assay and total cell yield was determined using an Invitrogen™ Countess™ Automated Cell Counter.
|Primary cortical neuron cultures at different developmental stages in culture. Mouse cortical neurons were isolated and cultured using the Thermo Scientific Pierce Primary Neuron Isolation Kit. Cortical neurons were grown on 24 well at the medium density of 1 x 10^5 cells or a 35mm glass bottom plate at the density of 5 x 10^6 cells. Upper panel: Phase-contrast images of mouse cerebral cortical cultures after 1, 14 and 28 days. After 1 day in culture, neurons start to differentiate and one or several short neurites extending from cell body can be observed; at Day 14 and Day 28, an extensive, intertwined network of axons and dendrites has developed. Lower panel: Immunostaining of cortical neurons at Day 1 and Day 14 with neuronal protein markers. Cells were fixed with 4% paraformaldehyde, permeabilized, and stained with microtubule-associated protein 2 (MAP2, green, Part No. 1861751), glial fibrillary acidic protein (GFAP, red), postsynaptic density protein 95 (PSD95, green), and synaptophysin (red). Images were taken at 20X, 40X, or 60X. The insert image indicates positive staining of synaptophysin and PSD95 at synaptic terminals, indicative of differentiated and functional neurons.
- Silva, R.F.M, et al. (2006) Dissociated primary nerve cell cultures as models for assessment of neurotoxicity. Toxicology Letters 163:1-9.
- Ivenshitz, M. and Segal, M, (2009). Neuronal density determines network connectivity and spontaneous activity in cultured hippocampus. J Neurophysiol 104:1052-60.
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