Several papers base their method of ES cell differentiation into neurons on Bibel et al., 2007 (Nat Prot). When the cells are initially plated for neuronal differentiation the medium contains N2 supplement and after a couple of days this is changed to B27 containing medium.
I'm wondering about the different effects N2 and B27 supplements have on differentiating neurons. Why is N2 used initially and then replaced by B27? Does N2 have any toxic effects on long-term cultures?
Hello Amila,
N2 supplement is used at initial stages because it helps in the initial commitment and differentiation, survival and expression of post mitotic neurons in culture. It favors the expression of neural specific genes plus it inhibits the growth of non neuronal cells in the culture (or any undifferentiated cells). That's why it is used during initial culture. Sometimes G5 supplement is also used in it's combination to increase the efficacy of neural differentiation. On the other hand B-27 supplement is used for long term survival of neurons and in addition to bFGF, it favors the growth and attachment of mature neurons.
Coming to the toxicity, N2 can be added to neuronal culture from initial to terminal phase of differentiation and same is true for B-27, but for B-27 initial attachment of cells is required. According to my experience, they didn't show any toxicity in long term cell culture, infect the combination helped to achieve a more potent neuronal phenotype.
Cheers !
what are the roles of B27,N2 in gall bladder organoid culture ?(non-neuronal)
N-2 Supplement is a chemically-defined, serum-free supplement based and recommended for growth and expression of neuroblastomas as well as post-mitotic neurons in primary cultures from both the peripheral nervous system and the central nervous system.
B-27 supplement improvements enable:
Increased neuronal survival by more than 50%—the highest in the industry
Accelerated neurite outgrowth
Masuma Akter - I might note that it is actually our newer B-27 Plus Supplement (TFS Cat# A3582801) and in combination with our newer Neurobasal Plus Medium, that we show an increase neuronal survival by more than 50% vs other systems, along with increase in neurite outgrowth and significantly improved electrophy activity. For these reasons, when culturing neurons we recommend trying B-27 Plus Supplement or the complete B-27 Plus Neuronal Culture system (Cat# A3653401) B-27 Plus + Neurobasal Plus) over our classic B-27 Supplement /Neurobasal system.
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