SeV is not pathogenic to humans (i.e., humans are not the natural host of the virus) and the infection does not persist in immunocompetent animals. Furthermore, SeV used in our kits does not produce infectious viral particles upon transduction to host hPSCs and is a temperature-sensitive mutant, such that it is active at 33℃ but becomes inactive at 37℃. However, because SeV can be transmitted by aerosol and contact with respiratory secretions and is highly contagious, appropriate care must be taken to prevent potential mucosal exposure to the virus. Our SeV-based kits must be used under Biosafety Level 2 (BL-2) containment with a biological safety cabinet or a laminar flow hood and with appropriate personal protective equipment. In the event that the virus comes into contact with skin or eyes, decontaminate the affected area by flushing with plenty of water and follow the safety manual prepared by your laboratory and approved by your Institutional Biosafety Committee.
Do I need a license agreement for any of Elixirgen Scientific’s products?
No. You don't need any licence or material transfer agreement (MTA) to use our differentiation kits or iPSC-derived cells. However, please be advised that these products are for research use only.
What kind of transcription factors are used for differentiation induction?
It is a proprietary formulated RNA and cannot be disclosed.
Does Quick-Tissue™ technology leave a genetic footprint?
Sendai virus (SeV) is an RNA virus, so it does not integrate into the genomic DNA. In principle, a foreign gene introduced intracellularly in the form of RNA is quickly translated and expressed because, unlike DNA, RNA does not need to enter the nucleus for forced expression, thereby providing no chance of mutagenesis. This is discussed in the following review paper: Yamamoto, et al., (2009) "Current prospects for mRNA gene delivery." Eur. J. Pharm Biopharm 71, 484-489.
User Guide Information/Troubleshooting
Why are there many aggregates after harvesting hPSCs?
After harvesting hPSCs, cells should not be left in the collection tube for more than 5 minutes. Leaving cells for a longer period may cause the formation of cellular clumps which interferes with infection. Single cells are best for infection.
I was able to harvest more hPSCs than I needed for the kit. Can I replate or freeze the leftover hPSCs to maintain their culture?
No. If you’re following the instructions given in the user guide, the hPSCs will be harvested in a differentiation medium so the leftover hPSCs cannot be maintained.
Can I use Geltrex or Matrigel as a coating substrate for hPSC cultures?
Our kits are not optimized with these substrates. However, if your hPSC differentiation protocols are already optimized with such a substrate, you may try using one of these with our kit at your own risk.
Can I use an orbital shaker for SeV infection while incubating cell suspension for 10 minutes?
The purpose of flicking the tube containing cells and SeV every 2 minutes during the 10 minute incubation is to prevent cells from settling down and forming aggregates. If the orbital shaker can do so, then it is possible to use it.