I am using DMEM media with 10% FBS and 1% Pen/Strp. It takes a long time for trypsinisation also. I grew the cells for a week and the cell count gave a count of 0.6 million cells only. I have attached the image of the cell after a week of incubation.
Were the cells thawed recently? Sometimes, it takes cells 2-3 passages before they really adopt their normal phenotype. Also: ATCC suggests splitting an 80% confluent plate 1:2, and replenishing medium every 3 days. This suggests, to me, that these are expected to be fairly slow growing cells in general. Further: some cells get really bad contact inhibition of growth, and when they grow in patches like that, they slow down a bit. Finally: I find that cells sometimes get resistant to trypsin if they've been left in the plate for too long.
Best recommendation: take that plate, split it 1:2, 1:4, and 1:8, and see what happens. Hopefully one of the plates will start growing more rapidly, and to greater confluence.
Yes, the cells were thawed recently.
Thank you for your reply. William P Katt I have split them into a T25 flask and am hopefully praying for the best.
Generally HT1376 cells are slow growing cells. Anyway try with higher percentage of FBS. Sometimes you need to provide little more nutrients to cells. therefor increase your FBS level up to 15 to 20%. I have recruited this method ,which works for me. Hope it will work.
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