I just started culturing NHEKs and I have no prior experience with them. I purchased them from Caltag (10HU-225) and using the PromoCell Serum-free Keratinocyte Growth Medium 2 (C-20011) with 0.06mM CaCl2 to keep them in the proliferation stage. What I'm seeing is that there's two distinct populations, with one being larger and often multi-nucleated, and the other one being smaller with more defined borders. Can someone provide any information on which population is normal and what the other cells are? I'm platting to do some ICC for early & late keratinote and fibroblast markers which hopefully will help. Thank you!
The mononucleated cells are your keratinocytes. The multinucleated cells, forming multinucleated giant cells, are derived from the keratinocytes under stress of various kinds, They are common in senescent cell cultures. In vivo, they form in skin subjected to extensive scratching, and also form in viral infection.
In vitro: Article Senescence-Associated Oxidative DNA Damage Promotes the Gene...
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