NS Supplement Series
NS supplement is a serum-free supplement for culturing neurons. It can be used to culture neurons and neural stem cells isolated from rat hippocampus. It should be used with NS basal medium and 200 mmol/L L-glutamine solution. Vitamin A-free and insulin-free products are also available.
Serum-free supplement for culturing neurons
NS supplement without insulin
NS supplement that does not contain insulin
Suitable for use in studying insulin secretion and receptors
NS supplement without vitamin A
NS supplement that does not contain vitamin A (retinyl acetate)
Vitamin A is considered to be involved in the differentiation of cells into neurons; thus, vitamin A-free reagents are used especially for culturing neural stem cells and glial cells.
Evaluation of NS Supplement
Cell number and expression of a neuron-specific marker for neurons derived from rat hippocampus
Figure 1: Comparison of the number of cells
Neurons were isolated from the hippocampus of rat embryo (E19), and were cultured on a plate coated with poly-L-lysine. NS supplement was added to NS basal medium to the final concentration of 2%. Neurons were cultured for 5 days and the number of cells were compared.
Green: neuron-specific marker (TuJ1) Blue: nucleus (DAPI)
Blue: nucleus (DAPI)
Figure 2: Expression of a neuron-specific marker
Cells were stained with a neuron-specific marker (TuJ1) and a nuclear marker (DAPI).
Evaluation of NS Supplement without insulin
Culturing primary neurons derived from rat cerebral cortex
Neurons were isolated from the cerebral cortex of rat embryo (E17), and were cultured in NS basal medium supplemented with either insulin-free or insulin-containing NS supplement (NSS). Cells were cultured for 6 days, and the number of cells grown with insulin-free supplement was compared relative to the number of cells grown with NSS (=1). As a comparison, cells were also cultured with insulin-free supplement containing 0.25x and 1.0x the amount of insulin in NSS.
NS basal medium + 2% NS supplement without insulin or NSS + 0.5 mmol/L L-glutamine (+ 0.25x or 1.0x insulin)
Number of cells seeded:
4.0×104 cells/well(96-well plate coated with poly-L-lysine (30-70 kDa)
Since insulin promotes survival of neurons, the number of viable cells was lower when they were grown with insulin-free NS supplement. However, with the addition of insulin, the number of cells was equivalent to those grown with NSS.
Evaluation of NS Supplements without vitamin A
Culturing neural stem cells derived from rat hippocampus
Neural stem cells were isolated from rat hippocampus, and were cultured in NS basal medium supplemented with either vitamin A-free or vitamin A-containing NS supplement (NSS). Cells were cultured for 3 days, and the number of cells grown with vitamin A-free supplement was compared relative to the number of cells grown with NSS (=1).
< Culture medium >
NS basal medium + 2% NS supplement without vitamin A, Competitor’s product, or NSS + 1 mmol/L L-glutamine
< Number of cells seeded >
12,000 cells/well (96-well plate coated with poly-L-lysine)
The numbers of cells grown with our NS supplement without vitamin A and the equivalent product from another company were comparable.
Culturing primary neurons derived from rat hippocampus
Neurons were isolated from the hippocampus of rat embryo (E19), and were cultured in NS basal medium supplemented with either our vitamin A-free NS supplement or an equivalent vitamin A-free product from another company. Cells were cultured for 6 days, and the expression of neuron-specific marker (β-III-tubulin, TuJ1) was examined.
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