Can I grow Gibco Rat Fetal Neural Stem Cells on cover slips?
If you are growing cells on cover slips to do immunocytochemistry, we recommend using Permanox chamber slides:
- Permanox chamber slides, 4-well, pk/16 (VWR, Cat. No. 62407-330)
- Permanox chamber slides, 8-well, pk/16 (VWR, Cat. No. 62407-335)
If you are growing cells on cover slips for other purposes and require coating, use this procedure to double coat the cover slips:
- Incubate cover slips overnight at room temperature in poly-ornithine solution (final concentration = 100 mg/mL in cell culture water)
- Rinse twice with sterile water
- Incubate cover slips overnight at 4 degrees C in laminin solution (final concentration = 15 mg/mL in cell culture water)
What are the estimated % neurons, astrocytes, and oligodendrocytes that we might get from using a spontaneous differentiation protocol vs. a directed differentiation protocol with Gibco Rat Fetal Neural Stem Cells?
Based on the expression of specific phenotypic markers, here are the expected percentages of the different cell populations:
Dcx marker (neuronal): 10% positive with spontaneous differentiation; 40% positive with directed differentiation
GFAP marker (astrocyte): 30% positive with spontaneous differentiation; 40% positive with directed differentiation
O4 marker (oligodendrocyte): 2% positive with spontaneous differentiation; 32% positive with directed differentiation
Is there any difference between Gibco Rat Fetal Neural Stem Cells isolated from cortex vs. hippocampus?
Cortical NSC's contain a greater mixture of cells and have different neurotransmitters. They have a wider set of applications. Fewer NSCs can be obtained from hippocampal tissue, but the cells are more pure. They are used by a smaller hippocampal-focused audience
Are Gibco Rat Fetal Neural Stem Cells derived from rats of a particular gender?
Gibco Rat Fetal Neural Stem Cells are derived from a mixed pool of embryos; therefore, cells will be derived from rats of both sexes.
What size container should I use for culture of Gibco Rat Fetal Neural Stem Cells?
Any format of culture vessel (e.g., flask, 6-well plates, etc.) can be used. Seed cells at a density of 5 x 10E4 cells/cm2. Depending on the surface area of your culture vessel, calculate how many cells you will need to have on hand. Example: For one 60 mm dish, you will need 5 x 10E4 cells/cm2 x 20 cm2 = 1 x 10E6 cells total.
Where can I find protocols for directed and spontaneous differentiation of Gibco Rat Fetal Neural Stem Cells?
You can find protocols for directed and spontaneous differentiation of Rat Neural Stem Cells in the Neurobiology Protocol Handbook on our website:
https://assets.thermofisher.com/TFS-Assets/BID/Handbooks/gibco-neurobiology-protocol-handbook.pdf
What are some of the key applications for Gibco Rat Fetal Neural Stem Cells?
Neural stem cells (NSC) is a valuable source not only for neuroscience but also for clinical use to treat neurodegenerative disease or neurological disorder. These cells can be used for a wide range of differentiation studies.
How do fetal derived NSCs compare with adult derived NSCs?
Adult derived NSCs tends to be regionally restricted and have limited plasticity compared to fetal derived NSCs.
Can I use Neurobasal medium/B27 or DMEM with Gibco Rat Fetal Neural Stem Cells instead of NSC-SFM?
To maintain the cells in NSC status, we recommend using NSC-SFM. We have validated and optimized the culture conditions in NSC-SFM to ensure superior growth and performance of these cells.
My Gibco Rat Fetal Neural Stem Cells do not attach to the CELLstart-coated plate but float to form a sphere. Is this normal?
Please ensure that the buffer used for CELLstart has Calcium and Magnesium. We recommend using Dulbecco's Phosphate Buffered Saline (D-PBS), containing calcium and magnesium, but no phenol red (Cat. No. 14040133). If the right buffer is used, try increasing the concentration of CELLstart up to 1:50 dilution rate. If CELLstart with our recommended buffer does not work, you can try using fibronectin or poly-L-ornithine-coated plates. If you use poly-L-orinithine, please perform an overnight incubation.
What is the doubling time for Gibco Rat Fetal Neural Stem Cells?
The doubling time for the cells is around 20-30 hrs. This tends to increase with passage number.
What viability should I expect from Gibco Rat Fetal Neural Stem Cells once thawed?
Upon thawing, the vial should have more than 50% viability which will provide more than 2 million live cells.
What is the % purity of Gibco Rat Fetal Neural Stem Cells?
Cell purity was measured by testing for the expression of the NSC phenotype marker, Nestin and the lack of expression of differentiated phenotype markers such as GFAP, Dcx, or GalC. More than 75% of the cells were positive for Nestin and expression of the differentiated markers was less than 10%.
How do you know that Gibco Rat Fetal Neural Stem Cells are indeed NSC?
We evaluated Gibco Rat Fetal Neural Stem Cells for two criteria:
- To express the NSC phenotype marker, Nestin and not express differentiated phenotype markers such as GFAP, Dcx, or GalC
- To demonstrate their differentiation potential toward all three downstream lineages such as neurons, astrocytes, and oligodendrocytes
For how many passages can I expand Gibco Rat Fetal Neural Stem Cells?
Rat NSC have been tested for proliferation and differentiation potential up to passage 3 after thawing and we have seen around 300 million cells from 1 vial after the 3rd passage. We have not expanded the cells beyond the third passage, but we encourage you to try expanding further.
At what passage were Gibco Rat Fetal Neural Stem Cells cryopreserved?
Cells were isolated from the fetus without passage and frozen down at passage 0.
Where were Gibco Rat Neural Stem Cells (Cat. No. N7744100) isolated from? How long can I culture these cells?
These cells were isolated from the cortex of Sprague-Dawley rats at day E14 of gestation. They can be expanded in culture up to 3 passages without differentiation.
What are the growth factors or cytokines used in neural stem cell (NSC) culture?
How can I culture human neural stem cells (NSCs)?
Human NSCs can grow in Gibco StemPro NSC SFM (Cat. No. A1050901) on dishes pre-coated with Gibco Geltrex Matrix or Gibco CELLstart substrate. Alternatively, if the goal is to obtain neurons, NSCs can also be grown on Neurobasal medium supplemented with Gibco B-27 supplements without vitamin A on a pre-coated dish.
How can I characterize neural stem cells (NSCs)?
NSCs are generally characterized by their ability to form neurospheres when plated at cloning density (Nat Methods 2:333 (2005)). NSCs can also be characterized by (1) RT-PCR of Sox1, Sox2, and Nestin or (2) immunohistochemical staining for nestin, Pax6, Sox2, and Ki67.
What are neural stem cells?
Neural stem cells (NSCs) are self-renewing multipotent cells of the nervous system capable of differentiating into neurons, oligodendrocytes, and astrocytes. NSC can be generated by induced differentiation from embryonic stem (ES) cells, or isolated from various regions of the brain including the cortex, the subventricular zone (SVZ), and the ventricular zone, or generated from bone marrow-derived mesenchymal stem cells (MSCs) (J Cell Biochem 114:764 (2013)). NSCs are valuable tools for the study of neurogenesis and neurotransmitter and receptor function. NSCs were used in the investigation of different CNS disorders such as PD and Huntington's disease in various animal models (J Cell Biochem 114:764 (2013)).
