1,25 Vit D3 reactivity - (Aug/17/2009 )

I'm trying to differentiate thp-1 with 1,25 vit-d3 and i was wondering how many days i can go without changing the media. if anybody has done this before: what did your differentiated cells look like and how many adherent cells did you get? My cells don't seem to be adhering.

help would be much appreciated!!!

thanks!

Hi,

It will depend on how much medium you add. But too much medium may prevent gas exchange if the culture is stationary. In principle, if the medium turns orange/yellowish fresh medium must be added. Or you could also add fresh medium w/D3 as time goes on, or split the cells. I use 100 nM D3 (Cayman) x 3 days without media change (3 million cells/50 ml RPMI), in my hands the cells keep proliferating and some will attach. I use them after 3 days.

good luck

V.