I am growing BT-483 in RPMI ATCC Mod + 20% FBS + 1% PSG + 0.01mg/mL bovine insulin and they are hardly growing in culture. I perform medium changes every 2-3 days. They take over 2 weeks to get to 80% confluent, and usually I split them before that because they grow upwards in plaques. The viability is also struggling and has been around 65-70% for the last 2 passages. They came out of thaw around 85%.
Any suggestions?
Hey Ashley, we're having the same issue. Tried new media (DMEM and 20% FBS) but still not much growth. Have you had any luck?
We found changing the basal medium actually ended up effecting the morphology over time (also tried DMEM in 20%).
We found a few things that helped get more consistent growth though. Seeding them around 6e6 in a T75 and about 1.5e7 in a T175 will be about 80-90% confluent in about 7-9 days. I also would recommend not doing a medium change after passaging for at least 2 days to allow as many cells to settle as possible. They're very sticky, so I would also recommend using your preferred dissociation reagent (tryple or trypsin) 2x after pre-rinsing the flask each time.
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