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| Catalog Number | Size | Price (USD) | Shopping Cart |
|---|---|---|---|
| NS36100 | 1 Million Cells | $295.00 | Buy Now | Add to Cart |
Neuroprogenitor Neurosphere tissue is provided live, unseparated, fresh from E18 rat cortex/hippocampus including subventricular zone. One order is from one brain which will provide over 1 million neurospheres. NNSph can be used for expansion as neurospheres with repeated passage or differentiation into neurons or astrocytes.
To optimize growth we provide: 12ml vial of Neurobasal/B27-retinoic acid/glutamax/egf/hgf (the last two are growth factors)- all components are also available through Invitrogen Corp.
Note: It is very important that these be cultured in Ultra Low Adhesion Wells. If they are put on a substrate then they will not create spheres. They must be cultured in suspension. See Corning Products, Options and Techniques.
Neurosphere FAQs:
1. Why do I see clusters forming right away?
Seeding cells at 30,000/cm2 will result in rapid cluster
formation. Without attachment, they will stay as progenitors
in Neuropro and grow into neurospheres that can be
harvested in 4-7 days. Evaluation with immunostaining for
nestin or other antibodies can begin after 4 days. To
see clonal growth, you can plate the original sample or these
neurospheres at limiting dilution of 50 cells/cm2 of tissue
culture plastic or ultra-low adhesion plastic in the same medium.
To see pluripotency, plate dissociated neurospheres at
5 to 15 thousand cells/cm2 on substrates coated with
poly-d-lysine (50 ug/ml water).
2. Should I seed single cells to do the neurosphere assay?
No, a neurosphere contains a cluster of cells. You can fix
them and stain as a cluster or dissociate them with papain
or trypsin, count to determine yield and/or fix to evaluate
individual cells.
3. The cells were single after seeding but they formed
cluster today. The sizes of some of the clusters
are very big. Is this the right sign?
Yes.
4. Should I observe the neurosphere formation to evaluate
the cells' progenitor stage?
No, wait at least 4 days.
5. How do I distinguish the neurosphere and cluster?
You can pass the neurospheres onto low adhesion plastic
or onto adhesive surface and see differentiation. As long
as cells are on low-adhesion plastic, they will stay as
progenitors.
6. Do I need to do immunostaining at this stage?
Wait at least 4 days.