NS21: Re-defined and modified supplement B27 for neuronal cultures

Department of Pharmacology, University of Iowa, Iowa City, IA 52242-1109, United States.
Journal of Neuroscience Methods (Impact Factor: 2.05). 07/2008; 171(2):239-47. DOI: 10.1016/j.jneumeth.2008.03.013
Source: PubMed


In vitro culturing of primary neurons is a mainstay of neurobiological research. Many of these culture paradigms have taken advantage of defined culture media rather than serum additives that contain undefined survival factors to facilitate experimental manipulations and interpretation of the results. To culture neurons in the absence of serum, defined supplements such as B27 are now widely used. However, commercially available supplements exhibit large variability in their capabilities to support neurons in culture. We re-optimized and modified earlier published formulations of B27 using 21 different ingredients (NS21). NS21 supports neuronal cultures of high quality as manifested by their morphological characteristics, formation of synapses, and postsynaptic responses. Much of the variability in the quality of B27/NS21 was due to variability in the quality of different sources of bovine serum albumin. Furthermore, we found that holo-transferrin used in NS21 is preferable over apo-transferrin used in B27 for the quality of neuronal cultures.

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    • "Some critical experiments of the distribution of synaptic proteins were analyzed using a blinding test to minimise the effects of biases. As dendritic spine formation is highly sensitive to culture conditions, such as the quality of B27 supplement (Chen et al., 2008), each set of experiments was usually repeated within a month using the same lot of B27 supplement and culture medium to reduce the variation. "
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    • "and include an array of amino acids and vitamins as well as traditional salts. Gem21 was used as previously described (Chen et al., 2008), and provided insulin, T3, and the antioxidant agents noted above. (Gemini Bio-Products Material Data Safety Sheet; www. "
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