Frequently Asked Questions

User Guide Information/Troubleshooting

How long do I need to maintain hPSCs prior to beginning to use your kit?

Before beginning differentiation, cultures should be healthy and displaying typical morphologies (i.e., round colonies with tightly packed cells exhibiting a small cytoplasmic area), few spontaneously differentiating cells, and normal growth rates (i.e., doubling once a day). We recommend growing the cells for at least 14 days post-thaw before differentiating and not allowing the cultures to reach more than 80% confluency.

Why do differentiation kits require hPSCs be maintained under StemFit or StemFlex conditions?

hPSCs should be adapted to passaging as single cells prior to differentiation. Our protocols have only been tested with StemFit/StemFlex, so we can't guarantee results using other culture conditions.

Why doesn’t Solution D1 dissociate my hPSC culture without scraping?

The concentration of the substrate used to coat the dish/well was likely too high, so Solution D1 treatment requires longer incubation than 10 minutes (up to 20 minutes). Next time, please reduce the concentration of the substrate to 50-70%.

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.