Instructions for use:
Prior to use, Neurobasal media must be supplemented with 0.5 mM L-glutamine for primary cells and 2.0 mM L-glutamine for neuronal phenotype tumor cells or neural stem cells. In addition, either a serum-free supplement or serum needs to be added. Recommended supplement is B-27 Supplement (Cat. No. 17504) for hippocampal and other CNS neurons. For embryonic neurons the addition of 25 uM glutamic acid is also recommended for the plating step. For neural stem cells N-2 Supplement (Cat. No. 17502) or B-27 Supplement without retinoic acid
(Cat. No. 12587) is recommended. For tumor cell lines of glial origin G-5 (Cat. No. 17503) is recommended.
NEUROBASAL Cat. No. 21103 - when supplemented with B27 is intended to give optimal growth and long- term survival of rat embryonic hippocampal neurons1, and growth and survival of neurons from embryonic rat striatum, substantia nigra, septum and cortex, and neonatal rat cerebellum and dentate gyrus2.
NEUROBASAL Cat. No. 12348 - Specific use for receptor studies such as estrogenic receptors, downstream protein purification studies or other processes where the presence of Phenol Red is undesirable
NEUROBASAL -A Cat. No. 10888 - when supplemented with B272 and β-FGF is intended to maintain long-term growth and viability of rat postnatal and adult hippocampal neurons.
NEUROBASAL -A Cat. No. 12349 - Specific use for receptor studies such as estrogenic receptors, downstream protein purification studies or other processes where the presence of Phenol Red is undesirable
NEUROBASAL with B27 (Cat. No. 17504) has shown excellent long-term viability of rat embryonic hippocampal neurons even after four (4) weeks in culture with greater than 90% viability for cells plated at 640/mm2 and
greater than 50% viability for cells plated at 160/mm2. Glial cell growth at five (5) days is reduced to less than 0.5% for a nearly pure neuronal population1 .
The growth of adult CNS neurons requires gentle separation of their numerous connections, a density gradient for the separation of oligodendrocytes and enrichment of neurons, an adequate substrate for attachment and a dedicated medium for growth. Brewer1 has shown that NEUROBASAL -A supplemented with B27 and adequate isolation methods permit the isolation of spherical remnants of hippocampal neurons from any age rat and promote the regeneration of axon and dendrite-like processes.
Neurobasal and Neurobasal -A must be supplemented with N2 or B27 and 0.5 mM L-glutamine. For the initial plating of embryonic primary hippocampal neurons, it is suggested that 25 μM (3.7 μg/mL) glutamate be added to the NEUROBASAL medium. Both media support the growth of nearly pure populations of neural cells without the need of an astrocyte feeder layer. Both media, when supplemented with B27, contain antioxidants to reduce reactive oxygen damage and they do not contain the excitatory amino acids, glutamate and aspartate, making it amenable to the study of these neurotransmitters.