My lab is starting a new line of iPSC culture (KOLF2.1j). The cell suppliers provide a maintenance culture protocol using StemFlex medium with RevitaCell Supplement (both Gibco) on Synthemax-II-SC Substrate (Corning) with ReLeSR (STEMCELL) for passaging. We currently maintain a different iPSC line (WTC11) in TeSR-E8 medium on Matrigel coating and use Accutase for passaging.
Do you think we will be able to use the TeSR-E8/matrigel combination for the new line? Or are different lines media/coat-specific? Will the cells be okay if I thaw them from StemFlex in to TeSR-E8? All these reagents are very expensive, and I would like to avoid buying them unnecessarily. I am the only person using iPSC cells here and I'm new to the process, so I can't ask anyone here
Dear Elizabeth Radley
i would not recommend it at all. The iPSCs do not handle this well (if you switch medium at the time of thawing). If you wish to adapt them I would thaw them in the same media that they were frozen in. Grow them for a passage or two and then start to adapt the cells to new media gradually.
Maybe ask another lab and borrow some small media volume from them if you wish to avoid buying the entire bottle. Although from my experience I would say it takes about a month or so to completely adapt the cells into new media and culture conditions.
good luck !
Hey Elizabeth Radley
I concur with Shambhabi Chatterjee .
I have done this many times when receiving iPSC from companies or other research groups. I will always revive with the media they used, with the ECM they use.
I would also recommend freezing down some wells using this combination as a backup.
I then switch one thing at a time. Usually the ECM first, if needed.
Then the media. I also try multiple wells with one being a complete switch to the new media, and others being various combinations of the old and new media.
Usually one of these wells looks good, and is handling it well, and you can transition to the new media slowly.
Chris
Hi! I am using the same line and had this same issue. I tried just thawing using the E8 media and matrigel and had poor cell survival so I thawing a new batch using the stemflex media and synthemax substrate.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies