Usually, basal media have been recommended for the culturing of human epidermal keratinocytes, and most articles suggest growing them in serum-free media. I'm finding it a bit challenging to understand proper cultivation methods for NHEK cells, can anyone please help me with this.
Hello Sabra Parveen ,
Assuming that the cells were newly acquired and primary, It would be wise to follow the culture media conditions recommended by the vendor. Beside the contents of the serum-free medium, the NHEK cells still need important factors and nutrients to grow.
Check out the recommended culture conditions from ATCC.
https://www.atcc.org/products/pcs-200-011
They recommend the serum-free basal medium + a few growth factors ( components of the keratinocytes growth kit)+ antibiotics (optional).
DMEM+FBS is often used to deactivate trysin during the passaging protocol
If you are isolating NHEK from scratch, check out this manuscript methods on page e6.
https://www.cell.com/cell-reports/pdf/S2211-1247(21)00002-4.pdf
These should reconcile some of the suggestions you see in articles.
Additionally, this inquiry thread may offer some insight in maintaining NHEK culture.
https://www.researchgate.net/post/Can-anyone-suggest-a-cell-culture-medium-for-normal-primary-Human-epithelial-keratinocyte-to-passage-them-for-more-than-7-passages
All the best!
Hello, mam.
That's quite helpful.
Thank you so much for your suggestions. 😊
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