|What is the difference between
Plus Supplement and classic
||B-27 Plus Supplement is an optimized formulation of classic
No small molecules or growth factors have been added.
|What is the difference between Neurobasal Plus Medium and Neurobasal Medium?
||Key amino acids and buffering components have been optimized in Neurobasal Plus Medium. No small molecules or growth factors have been added.
|How did you upgrade the manufacturing/QC process?
||We streamlined the manufacturing process, implemented more stringent acceptance criteria and enhanced QC assays for qualification of raw materials and release of finished product to enable better lot to lot consistency.
|Can I re-freeze B-27 Plus into smaller aliquots for future use?
||Yes, we recommend aliquoting upon first thaw. We recommend no more than 2 total freeze/thaw cycles.
|Can I use the B-27 Plus Neuronal Culture System for expansion of proliferating cells, such as neural stem cells?
||No, we do not recommend the B-27 Plus System for expanding proliferating cells. We recommend using StemPro NSC SFM, Neural Induction Medium Supplement or B-27 Supplement minus Vitamin A for growing proliferative cells, such as NSCs.
|How long can I maintain my neurons in culture with the B-27 Plus System?
||We have maintained primary rat cortical neurons up to 8 weeks and rat hippocampal neurons up to 4 weeks. Mouse cortical neurons were maintained for 4 weeks. Human iPSC-derived neurons (HIP™ Neurons from MTI-GlobalStem) have been maintained for 5 weeks. All cells tested with the B-27 Plus System were significantly healthier than the classic products across all time points.
|Do I have to use B-27 Plus with Neurobasal Plus?
||B-27 Plus Supplement and Neurobasal Plus Medium have been designed to work synergistically together for maximal performance. They can be used with other basal media or supplements if necessary.
|How can I control the number of glial cells in my culture?
||Add CultureOne Supplement at day 0 with the B-27 Plus Neuronal Culture System to fully suppress both astrocytes and oligodendrocytes with no detrimental effect on neurons. Delaying the addition of CultureOne to later time points results in increasing levels of astrocytes. Refer to the CultureOne Supplement user guide for more information.
|Can I use B-27 Plus Neuronal Culture System to culture non-neuronal cells?
||We have not tested in other cells, however B-27 Plus System was designed for culturing neuronal cells.
|Which classic B-27 and Neurobasal products can I substitute B-27 Plus and Neurobasal Plus for?
||Substitute for the regular versions of B-27 Supplement and Neurobasal Medium products (not specialty versions like minus Insulin, minus Vitamin-A or Neurobasal-A) in protocols used for:
- Maintenance of primary rat, mouse and human PSC-derived and fetal-derived neurons
- Differentiation of human PSC-derived and fetal-derived Neural Stem Cells (NSC) to neurons
|How long can I use complete Neurobasal Plus supplemented media?
||Complete Neurobasal Plus Medium should be used within 4 weeks after supplementation with B-27 Plus Supplement.
|Can thawed B-27 Plus Supplement be kept at 4C?
||Yes, B-27 Plus Supplement can be stored at 4C for up to 2 weeks post thaw.
|What is the shelf-life of B-27 Plus Supplement and Neurobasal Plus Medium?
|What do I do if my cells are not attaching or are clumped?
||In primary neuronal cultures this is typically a function of uneven coating or if neurons are plated at too high of a density. In NSC differentiation experiments, this could be due to over proliferation of neural progenitor cells. Differentiating with the B-27 Plus Neuronal Culture System and CultureOne Supplement can eliminate more than 75% of neural progenitor cells and the clumps they form. See CultureOne Supplement for more information.
|What is HuC/D?
||HuC/D is an antibody specific to neuronal cell bodies. It can be used to quantitate neurons in culture.
|How do I thaw B-27 Plus Supplement?
||Thaw at 4C overnight or at room temperature for 2 hours. We don't recommend thawing in a water bath or incubator.
|What happens if I thaw B-27 Plus Supplement in a 37C water bath or incubator?
||This is not recommended.
|How do I feed my cells with the B-27 Plus Neuronal Culture System?
||Perform half medium exchanges every 2-3 days post plating. Make sure that neurons are not completely exposed to air by not removing all of the medium during exchanges.