I'm using cell to produce Wnt-3a conditioned medium to growth organoid. How can I measure the activity of the medium ? I also would like to check the performance of my cells . Do they have limited passages ?
Hello
Look at these links
Hope help you
http://www.bio-protocol.org/e1183
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4401205/
Hi Benedetta,
The protocol Saleh describes works perfectly for me. You can really speed up the protocol however. I personally transfect while plating into a 96-well. This will make sure you have equal transfection per well. Change medium the day after and you can harvest already the same day if you do it early in the morning. Otherwise 24h later. This protocol is therefore for 48h instead of 5 days.
Your Wnt CM should give at least 5x higher results when compared to standard medium if using 1:1 ratio (Wnt:DMEM). I never experienced higher results than 50x. If you want you can even test your RSPO CM. I usually mix those 1:1:1. This should add another max up to 200x in comparison to your Wnt-only condition.
Hope this helps
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