Help me....About the 3T3-L1 Cells - (Sep/11/2005 )
As you know, form planting to inducing differentiation and
maintaining for differentiation of 3T3-L1 preadipocyte cells, we need
in all about 6~8 days. We did as the most classic references said,
inducing differentiation at the 0 day of the confluence, but
unfortunately, after 2 days of inducing, the 3T3-L1 cells were easily
scratch from the well as we replenished the culture media. So that was
blocked me of my further investigation.
Can you give me some advice? What can we do to make the 3T3-L1 cells
adhere to the bottom of the well firmly?
inducing differentiation at the 0 day of the confluence, but
unfortunately, after 2 days of inducing, the 3T3-L1 cells were easily
scratch from the well as we replenished the culture media.
Hello,
Ussually, after induced 2 days, cells are so sensitive so u need to do carefully whenever u want to change with fresh media. It's better that using pipette man and gentlely drawing old media from the well and then also slowly adding new culture media.
Try!!! And good luck to you.
We usually induce differentiation (i.e. give the differentiation cocktail) 48 hours after the confluence.
Anyway, since that moment on, cells start to produce lipid droplets that make them less tight and, it's difficult to explain, more delicate to the medium flow when you add it through the pipette.
So take 2 minutes more, but add the medium very very slowly.