Breadcrumb
- Home
- Products
- Neural Stem Cells
- Neural Stem Cells Cell Culture Products
- N21-MAX Media Supplement (50X) (AR008)
N21-MAX Media Supplement (50X) Best Seller
R&D Systems, part of Bio-Techne | Catalog # AR008
Key Product Details
Summary for N21-MAX Media Supplement (50X)
For the reliable maturation and superior long-term culture of primary neurons.
Key Benefits
- Provides reliable maturation and superior function of neurons in culture
- Can be used to improve stem cell and 3D culture conditions
- Rigorous quality control testing ensures consistency across experiments
- Is nutrient-rich and serum-free
- Eliminates the need for an astrocyte feeder layer in neuronal cultures
Why Culture Neurons in Serum-Free N21-MAX Media Supplement?
The N21-MAX Media Supplement improves upon the traditional B271 and NS212 neuronal supplements, offering a serum-free and fully-defined formulation that is optimized for the reliable maturation, consistent health, and superior function of neurons in culture. There is an increase in the use of B27-like supplements to optimize stem cell and 3D cell culture media. N21-MAX improves stem cell differentiation and increases the viability and health of differentiated cell types during long-term cell culture conditions.
N21-MAX was designed to eliminate uncontrolled variables, such as those found in both serum-containing media and in some commercial serum-free neuronal supplements. Use of undefined media supplements can confound neuronal growth and maturation. N21-MAX contains a cocktail of factors optimized for neuronal survival, neurite extension, and synaptic maturation. Each lot of N21-MAX is tested on primary hippocampal neurons to ensure consistent performance for the user.
The N21-MAX Media Supplement:
- Is fully defined and quality tested to reduce unwanted experimental variation.
- Contains factors qualified to support reproducible and long-term neuron culture.
- Modified from the published B27 formulation1.
- Does not require an astrocyte feeder layer.
- Has been tested to support the viability and growth of E18 rat hippocampal neurons.
- Brewer et al., (1993) J. Neurosci. Res. 32:567
- Chen et al., (2008) J. Neurosci. Methods 171:239
Kit Contents
N21-MAX is optimized for the maturation and long-term culturing of neuronal cells. The supplement is supplied as a 50X concentrated solution and contains the following 21 components:
- Albumin (bovine)
- L-Carnitine
- Catalase
- Corticosterone
- Ethanolamine
- Glutathione
- Galactose
- Holo-Transferrin
- Insulin
- Linoleic Acid
- Linolenic Acid
- Lipoic Acid
- Progesterone
- Putrescine
- Retinyl acetate
- Retinol
- Selenite
- Superoxide dismutase
- Triiodo-L-thyronine
- D,L-alpha-Tocopherol
- D,L-alpha-Tocopherol acetate
Supplied in a volume sufficient to supplement 500 mL of media at the recommended concentration. N21-MAX is tested for use in Neurobasal Media.
Precautions
This product contains human transferrin. This transferrin was purified from donor plasma and 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.
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.
Source | N/A |
|
N21-MAX Reduces Spontaneous Stress Rod Formation. E18 rat hippocampal neuron cultures were plated on poly-D-lysine coated coverslips and grown for 5 days in media supplement with either N21-MAX (Catalog # AR008) or another commercial neural media supplement (Competitor). Cells were fixed and immunostained for rod formation, and indicator of cellular stress, using an antibody against cofilin (% of Neurons with Rods). A lower percentage of hippocampal neurons that were cultured in N21-MAX had spontaneous cofilin rod formation (4.5%) compared to neurons cultured in competitor media (16.8%). Data courtesy of the laboratory of Dr. James Bamburg at Colorado State University. |
|
Improved Resolution of Rod Induction in Neurons Cultured in N21-MAX. E18 rat hippocampal neuron cultures were plated on poly-D-lysine coated coverslips and grown for 5 days in N21-MAX (Catalog # AR008). Neurons were then treated with rod-inducing agents or maintained in N21-MAX alone (Untreated). Neurons were fixed and immunostained for rod formation using an antibody against cofilin (% of Neurons with Rods). The low background of rod formation in untreated cultures enabled detection of mild rod induction using Synthetic Amyloid beta, Amyloid beta dimer/trimer, and TNF-alpha. As a positive control, treatment with an ATP depletion solution or glutamate resulted in robust rod induction. Data courtesy of the laboratory of Dr. James Bamburg at Colorado State University. |
|
N21-MAX Media Supplement Enhances Synaptic Development. E18 rat hippocampal neurons were grown for 19 days in vitro in media supplemented with either N21-MAX Media Supplement or the neural media supplement from the most widely-used competitor. Cells were incubated with the synaptic vesicle dye, SynaptoRed™C2 (10 µM; Catalog # 5118), for 1 minute prior to depolarization with 50 mM KCl. Cells were imaged for SynaptoRed™C2-positive synaptic puncta using the Operetta CLS™High Content Analysis System. (A) Quantification of SynaptoRed™C2-positive puncta shows that neurons grown in N21-MAX have an increased number of synaptic puncta compared to the competitor media. (B) Quantification of dye intensity shows that neurons grown in N21-MAX have more robust synaptic activity than neurons cultured in the competitor media. (C) Representative images of SynaptoRed™-C2 staining in neurons cultured in N21-MAX or competitor media. (D) Representative images showing the quantification of synaptic puncta in neurons cultured in N21-MAX or competitor media. Colored circles indicate a SynaptoRed™-positive synaptic puncta. |
|
Synaptotagmin and CAM Kinase II Expression in Neurons Cultured in Media Supplemented with N21-MAX. Synaptotagmin and CAM Kinase II Expression in Neurons Cultured in Media Supplemented with N21-MAX.E18 rat hippocampal neurons were grown for 21 daysin vitroin media supplemented with N21-MAX Media Supplement (Catalog # AR008). Cells were stained with either (A) a Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364), followed by the NorthernLights™(NL)557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; yellow), or (B) a Mouse Anti-Human CaM Kinase II alpha Monoclonal Antibody (Catalog # MAB5584), followed by the NL557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; green). The nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips. |
|
Increased Synaptic Puncta and Neurite Outgrowth of Primary Neurons Cultured in N21-MAX. Increased Synaptic Puncta and Neurite Outgrowth of Primary Neurons Cultured in N21-MAX.E18 rat hippocampal neurons were grown for 21 daysin vitroin media supplemented with either N21-MAX Media Supplement (Catalog # AR008) or the neural media supplement from the most widely-used competitor. Staining for Synaptotagmin (yellow) showed more robust synaptic puncta and increased neurite outgrowth in neurons cultured in N21-MAX compared to those cultured in competitor media. Cells were staineed with a Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) followed by the NorthernLights™(NL)557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007). Nuclei were counterstained with DAPI (blue). |
|
N21-MAX Media Supplement Increases Efficiency of Pancreatic Cell Differentiation. N21-MAX Media Supplement Increases Efficiency of Pancreatic Cell Differentiation.Differentiation of PDX-1+pancreatic cells from induced pluripotent stem cells (iPS2) was performed with base media containing either the N21-MAX Media Supplement (green) or the competitor equivalent (red).A) Compared to competitor media, N21-MAX improved pancreatic cell differentiation as observed by flow cytometry.B) Bar graph quantifying PDX-1+cells in N21-MAX and the competitor media. |
Preparation & Storage
Shipping Conditions | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
Storage | Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date. |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, completed N21-MAX-supplemented medium is prepared using the following procedure:
- Dilute the media supplement in basal media
- Store completed media
- Use within 2 weeks
Reagents Provided
Reagents provided in the N21-MAX Media Supplement (Catalog # AR008):
- Albumin (bovine)
- L-Carnitine
- Catalase
- Corticosterone
- Ethanolamine
- Glutathione
- Galactose
- Holo-Transferrin
- Insulin
- Linoleic Acid
- Linolenic Acid
- Lipoic Acid
- Progesterone
- Putrescine
- Retinyl acetate
- Retinol
- Selenite
- Superoxide dismutase
- Triiodo-L-thyronine
- D,L-alpha-Tocopherol
- D,L-alpha-Tocopherol acetate
Other Supplies Required
Reagents
- Basal media (e.g., Neurobasal Media from Invitrogen®, Catalog # 21103-029 or equivalent)
- L-Glutamine
Materials
- Serological pipettes
- Pipettes and pipette tips
Equipment
- 2 °C to 8 °C refrigerator
Protocol Overview
Dilute 50-fold with a basal media (e.g., Neurobasal Media from Invitrogen, Catalog # 21103-029) and supplement with 0.5 mM L-glutamine before use. The medium may be stored in the dark at 2 °C to 8 °C for up to 2 weeks.
Invitrogen is a registered trademark of Invitrogen Corp.