Hello Guys.
I am culturing adult Human Endothelial Keratinocyte (HEK) cells and a recent batch have got some unusual morphology. Instead of a uniform cobblestone appearance , I am observing additional elongated cells as well as stringy structures. The monolayer generally is looking quite messy and uneven.
I am attaching one image from the previous batches, which is a uniform cobblestone monolayer, and some pictures of the new batch.
The cells are all primary and were purchased from the same supplier. The passages of the cells are before the senescence P3 (senescence comes around P7).
The composition of the media, the cells are cultured in, is the same.
Thank you in advance
Yegor
The elongated cells look like fibroblasts, which normally end up within a keratinocyte culture but apparently failed to be removed from this batch. They easily are removed, as they are much more sensitive to trypsin (0.05% is sufficient) release from the plate. 5 to 10 minutes should be adequate to remove them and the keratinocytes should be unaffected as long as you wash gently and don't leave the trypsin on too long.
You're welcome. I also just thought of a second way to correct this...
Since these were purchased, contact the vendor and tell them to send you a better batch! At a minimum they should offer you a discount on a future purchase given that they failed to provide you with a pure cell population.
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