Hey all
I tried reviving the HaCaT cell line using the standard protocol for cell revival. However, they aren't attached to the flask even after 72 hours. The cell lines were stored in August 2022 by a colleague of mine. What would be the reason behind their failure to revive?
Media used: DMEM HG 12% FBS
The recommended media by ATCC for culturing HaCaT is Dermal Cell Basal Medium (https://www.atcc.org/products/pcs-200-011#required-products). However, our lab has previously grown and cultivated them in DMEM with 12% FBS. Others have used DMEM with 5% FBS. (Article Insulin–cobalt core–shell nanoparticles for receptor-targete...
Could the problem be due to the difference in media? Or the stress due to storage conditions that prevented its revival?
Are these HaCaT cells stored in -80 really lost?
Please have a look at the image.
Thank you
These cells are relatively difficult to culture, and tend to age (differentiate), gradually become thorns (with many thorns), and then become small pellets (when the cells generally like to be tightly clustered together), and finally fall off into suspended cells. Which DMEM HG medium did you use? https://www.thermofisher.cn/cn/zh/home/technical-resources/media-formulation.8.html. This is the calcium-containing one, and the calcium-free one can slow down the rate of differentiation, and of course some people add NEAA
Thank you for your reply Ke Zhao . I used https://www.himedialabs.com/in/al007a-dulbeccos-modified-eagle-medium-dmem-high-glucose-w-4-5gms-glucose-per-litre-l-glutamine-3-7-gms-per-litre-sodium-bicarbonate-and-sodium-pyruvate.html from Hi-Media. It doesn't have Calcium. Is calcium essential for this cell line?
- Badges
- Science method