Although there are a number of protocols available to isolate rat cortical stem cells, each isolation can yield cells that vary in yield, purity, and their ability to proliferate. See Details
R&D Systems offers Rat Cortical Stem Cells that are of high purity, frozen at a Passage 0, and are validated to be free of mycoplasma and microbial contamination. Rat primary cortical stem cells were isolated from the cortex of embryonic E14.5 Sprague-Dawley rats. The cells were cultured in a monolayer system in medium supplemented with N-2 Plus Media Supplement (Catalog # AR003) or N-2 MAX Media Supplement (Catalog # AR009) and Recombinant Human FGF basic (Catalog # 233-FB or 4114-TC). Cells were then harvested and cryopreserved. The cells are designated as passage 0 (P0) cells.
R&D Systems Rat Cortical Stem Cells:
1 vial of Rat Cortical Stem Cells containing 3 x 106 cells. See Details
Store in liquid nitrogen for up to 1 year.
This product contains trace amounts of human transferrin and DMSO. The transferrin was tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen. As no testing can offer complete assurance of freedom from infectious agents, these reagents should be handled as if capable of transmitting infection.
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Differentiation of Rat Cortical Stem Cells. Neural progenitors were cultured for seven days in DMEM/F12 containing N-2 Plus Media Supplement (Catalog # AR003) without FGF Basic to induce differentiation. The differentiated cells were labeled with a Goat Anti-Rat Nestin Affinity-purified Polyclonal Antibody (Catalog # AF2736) followed by the NorthernLights™(NL)493-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL003; green) and a Mouse Neuron-specific Anti-beta-III Tubulin Monoclonal Antibody (Catalog # MAB1195) followed by the NL-557-conjugated Donkey IgG Anti-Mouse Secondary Antibody (Catalog # NL007; red). Nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
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Neural Progenitor Cells Expanded with N-2 Plus Media Supplement Express Nestin and SOX2. Rat Cortical Stem Cells (Catalog # NSC001) were cultured for 7 days in media supplemented with 1X N-2 Plus Media Supplement (Catalog #AR003) and 20 ng/mL of Recombinant Human FGF basic (Catalog # 233-FB). The cells were stained with a PE-conjugated Mouse Anti-Human Nestin Monoclonal Antibody (Catalog # IC1259P, red histogram), a PE-conjugated Mouse Anti-Human/Mouse SOX2 Monoclonal Antibody (Catalog # IC2018P, green histogram), or a PE-conjugated Mouse IgG2A Isotype Control Antibody (Catalog # IC003P, open histogram). Under these conditions, cells were shown to express high levels of neural multipotency markers Nestin and SOX2.
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Verification of Neural Progenitor Cell Multipotency Following Expansion with N-2 MAX Media Supplement. Rat Cortical Stem Cells (Catalog # NSC001) were differentiated for 7 days in media supplemented with N-2 MAX Media Supplement (Catalog # AR009). Differentiated cells were stained with a Mouse Neuron-specific Anti-beta-III Tubulin Monoclonal Antibody (Catalog # MAB1195) followed by the NorthernLights™ (NL)557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; red) to detect neurons. The cells were stained with a Sheep Anti-Human/Rat GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594) followed by the NL557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) to detect astrocytes; and cells were stained with a Mouse Anti-Human/Mouse/Rat/Chicken Oligodendrocyte Marker O4 Monoclonal Antibody (Catalog # MAB1326), followed by the NL 557-conjugated Goat Anti-Mouse IgM Secondary Antibody (Catalog # NL019) to detect oligodendrocytes. The nuclei were counterstained with DAPI (blue).
Neural stem cells provide an excellent model for research focused on neural development and neurological disorders. R&D Systems offers ready-to-use primary cortical stem cells isolated from E14.5 Sprague-Dawley rats. In addition, primary mouse cortical stem cells isolated from E14.5 CD-1 mice are available. Every lot of R&D Systems Cortical Stem Cells is validated for a high level of Nestin expression and the capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes. Our cortical stem cells are tested to ensure highest quality and lot to lot consistency. Both rat and mouse Cortical Stem Cells can be optimally expanded as monolayers or neurospheres.
To complement the use of primary neural stem cells, we offer a range of supportive products, including culture media which is specifically optimized for use with neural stem cells. We offer kits to promote the in vitro proliferation of neural precursors, and kits to differentiate neural stem cells into dopaminergic neurons or oligodendrocytes. In addition, kits are available which contain panels of antibodies designed to monitor the differentiation and identification of neural precursors, astrocytes, neurons, and oligodendrocytes.
Refer to the product datasheet for complete product details.
Reagents Supplied in Rat Cortical Stem Cells (Catalog # NSC001)
Expanding Rat Cortical Stem Cells Using the Neurosphere System
Expanding Rat Cortical Stem Cells Using the Monolayer System
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