10/21/2011 9:35:00 AM
Description:
The STEMdiff™ Neural Induction Culture System is a reliable, flexible and complete system for the efficient and reproducible derivation of neural progenitor cells (NPCs) from human pluripotent stem cells (hPSCs). The media are optimized and defined for consistent results, and the procedure is fast and convenient compared to other methods of neural induction.
STEMdiff™ Neural Induction Medium (Catalog #05831) allows for the highly efficient differentiation of hPSCs to neural rosettes, and
STEMdiff™ Neural Rosette Selection Reagent (Catalog # 05832) selectively detaches neural rosettes for subsequent isolation of NPCs at high purity.
Optimizing the procedure from beginning to end gives superior results! Here are some important tips for successful neural induction:
- If starting with hPSC grown on feeders, it may be best to switch to a feeder-independent system using mTeSR®1 (Catalog # 05850) plus hESC-qualified BD Matrigel™ (BD Catalog # 354277) or StemAdhere™ (Catalog # 07170) first before attempting the neural induction protocol.
- It is important to start with a high quality culture. This means that no more than 10% of the colonies in the starting population should display morphological differentiation, and cells should be in growth phase.
- Use a ‘control’ cell line the first time you use this protocol. The following cell lines have been tested and found to work using the protocol provided in our Technical Manual: H1, H7, H9 and iPSC-4D1
- The protocol may need some adapting due to inherent variations between all available hESC lines and iPSC lines.
- Use the AggreWell™800 (Catalog #27865/27965) plate, as larger aggregates are more efficient in terms of NPC isolation later on.
- To avoid co-selection of flat cells (neural crest cells), do not over-incubate with STEMdiff™ Neural Rosette Selection Reagent or excessively wash the plate with DMEM/F12 when removing the neural rosette clusters.
- Pre-warm the media before use; do not add cold media from the fridge directly to your cells.
- Confirm neural rosette formation by immunocytochemistry:
| Marker |
Cell Type |
| Pax-6 |
Early neural progenitor |
| ZO-1 |
Rosette Lumen |
| Sox-1 |
Neural progenitor cells |
| Nestin |
Pan-neural |
| Sox-10 |
Flat, neural crest |
NPCs derived using our STEMdiff™ Neural Induction Culture System are shown to be functional and capable of differentiation to mature neuronal and glial cells. For more information on the downstream applications for NPCs derived using this method, please refer to the
Technical Bulletin.
For further assistance and information, please contact
techsupport@stemcell.com.