NeuroMax (B27) Supplement, 50-x solution, sterile
NeuroMax is a serum free chemically defined supplement for long-term viability and growth of hippocampal and other neurons of the central and peripheral nervous system. NeuroMax supports the growth of neuronal cells without the need of specific feeder cells. It contains vitamins, antioxidants, hormones, lipids, albumin, insulin, transferrin.
Field of application: neurobiology, biomedical research.
Recommended media framework for optimization: Neurobasal (Neurobasal™ Medium).
This product is intended for research purposes.
Supported cell cultures of neuronal origin:
- Embryonic and prenatal neuroblasts and progenitors.
- Neurons of adult nervous tissue.
- Primary cultures of tumor cells and permanent cell lines of tumor tissues of the nervous system.
- Embryonic neurons of the striatum and the rat midbrain (EGF-stimulated growth progression).
PREPARATION INSTRUCTIONS:
One bottle of the product (10 ml) is enough for 500 ml of optimized media.
Before use, B27 should be thawed. Defrosting should occur naturally in the refrigerator at temperature of +2 to +8 °C.
The described testing procedure was used to cultivate neurons of hippocampus and cerebral cortex of an 18-day-old rat embryo, as well as neuroblastoma cell lines.
- Cover the surface of culture container with adhesive: sterile aqueous solution of poly-D-lysine 0.05 mg/ml at the rate of 0.15 ml/cm² (withstand 1 hour at room temperature).
- Remove the adhesive solution and rinse the surface twice with sterile deionized water. Completely dry the containers in a laminar flow cabinet. Use immediately or store in the refrigerator for up to 2 weeks.
CULTIVATION METHOD:
- Isolate the primary neurons of a rat or use a culture of frozen neurons according to standard laboratory protocols.
- Seeding the cells in heated at 37 °C full Neurobasal media. The recommended passage density is ~160 cells/cm².
- Incubate vessels at 37 °C under standard conditions of a CO₂ incubator, however, it is preferable to reduce the oxygen to 9%.
- After 4-24 hours, replace ½ of the media for fresh one.
In case of non-hippocampal culture of neurons: 4 days after the passage, replace ½ of the media for fresh one. For hippocampal culture of neurons: 3 days after the passage, replace ½ of the media for fresh one without L-glutamate. Repeat the procedure every 3 days. (Addition of 25 microns of 2-mercaptoethanol increases the survival rate and duration of cultivation of hippocampal neurons in vitro).
HOW SUPPLIED: in sterile plastic bottles of 10 ml.
SHELF LIFE: 2 years.
STORAGE CONDITIONS: -10 to -20 °C in a place protected from light.
RECOMMENDED USE:
- Do not use the product after its expiry date.
- Use gloves, mask and clothes adapted to the manipulation of the product to avoid contamination.
- Manipulate the product in aseptic conditions.
- Supplements such as antibiotics or L-Glutamine should be added as sterile supplements to the ready medium.
- Solution should be clear and free of opalescence and sediment.
