Discoloration in cell culture conditions
Lately, Anant and I have been doing our discoloration experiments under purposely harsh conditions so we don’t have to wait for multiple days for experiments to finish.
We have lots of evidence that the 3D format slows down discoloration (post1, post2, post3). This experiment was to test how long samples will last in 3D format under real cell culture conditions. These samples were from w413. Dave RTP’ed them for me (800C, 5min, Ar). Then I assembled them (or not) into Sepcons and then autoclaved (or not). Cell conditions were: incubator, DMEM+10%FBS.
The 2D format was control. As expected, the RTP treatment slowed discoloration from 1 day to 2 days. Autoclaving untreated samples slowed discoloration here, and in this experiment, autoclaving after RTP further inhibited discoloration by ~ 1 day. This is the longest a sample has lasted in DMEM in the incubator in a long time (in 2D format).
Simply putting these samples in the transwell format doubled the time to discoloration, regardless of the treatment. So RTPing and then autoclaving (which needs to be done for cell culture anyway) the pnc-Si permits almost 1 week of stability under cell culture conditions in DMEM. We don’t even use DMEM anymore since we’re mainly working with HUVEC, so it’s likely that we can expect at least 1 week before discoloration in 3D format.