Hello everyone, I am working with 3t3-l1 cell and I have been having a problem with peeling and curling effect when differentiated. My cells start to peel off and curl one or two days after changing to maturation media. However, this problem is not occur all times. Sometimes it differentiates well but sometimes it peels off. From my searching to find out any solutions, I think this problem might due to passage effect because the cell first I got from other lab is from Atcc source and passage greater than 20. So, I want to buy new one. However, I still hesitate to choose between 3t3-l1 from zen-bio or ATCC. Are there any suggestions? Which one gives better result for differentiation?
Thank you in advance
Ameena.
As 3T3-L1 is not only available from ATCC and ZenBio, but also from ECACC, BCRC, BCRJ, JCRB/IFO, KCLB, Kerafast and probably others:
http://web.expasy.org/cellosaurus/CVCL_0123
you should maybe "widen" the question to ask about the experience of other researchers with various batches of 3T3-L1.
Best
Amos
Hello
zen-bio, and use with it this protocol
Good Luck
https://www.biocat.com/bc/pdf/ZenBio_3T3_L1_User_Manual.pdf
Hello,
I highly recommend you read the following article. I think it would be extremely helpful.
Best,
-Matt
http://www.nature.com/articles/srep42104
Thanks, Matt! The article he linked above covers some of the variances between 3T3-L1 cells from various sources. I would certainly not purchase them from ECACC, nor would I from ATCC - they are maintaining 5 different lots right now with varying degrees of differentiation ability. Zenbio cells are sourced directly from the origin lab at Harvard, so you know you're getting exactly those - unlike with whatever the ATCC or other suppliers are offering. Which the literature would suggest is extremely variable.
To your problem though, what type of cell culture dishes are you using? I've found that 12 and 24-well plates have this issue more than when I culture in 96-well plates. Ultimately, in my experience, this is less about passage number and more about cell handling. You have to be extremely careful when removing and adding back fresh media to avoid disrupting the edge and getting this folding. I've done cell work for many years, but had to practice with this to get the technique just right. As in, visually inspect the cells under the microscope, ensure there is no issue, then remove your media and add it back from a single row. Then visually re-inspect that row. Ensure you've not disturbed them at all. If you have, try to adjust your technique slightly for the next row. They are fragile cells, so I exaggerate the care with which I remove and add my media back. Slow and steady!
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