I dont know what is Incap cells, do you mind explaining?
I have similar problem with MCF-7. I start to notice that MCF-7 is might be sensitive to ethanol. I made it custom to spray ethanol on the body of the flasks everytime removing or keeping it in the incubator. I try not to spray the caps. I notice that my media gets cloudy in 24h as the cells lysed. A PHD student recommend me not to spray anymore, when I stop spraying, the condition get so much better, MCF7 becomes easily confluent. Why?
I posted my question as new topic just now, maybe you could give some of your opinion.
In some published paper, some people use 0.1%ethanol as vehicle control,but my 0.1%ethanol group were all dead, but 0.1%DMSO group was OK, maybe different resource, our MCF-7 cells were just sensitive to ethanol, now, I dissolved all my reagents in DMSO, my experiment is going on well. Good luck to you.
I have been culturing cells more than one year, cultured around 20 kinds of cells, but recently, I found my lncap cells and MCF-7 cells are ethanol-sensitive, when treated with 0.1% ethanol, they all died, but 0.01%ethanol group is OK, 0.05% group, half of the cells died. I repeated 2-3 times, the same thing happened. but, years ago, there was another person doing experiments with lncap cells in our lab, she used 0.2% ethanol, the cells were still OK(both my cells and her cells are from the same original vial ordered from ATCC), I don't know what happened to these cells? Does anyone have this kind of experience?
PS: At the same time, I am using LAPC-4, HAEC cells doing experients, they are OK when using 0.1% ethanol.
I need to do cell proliferation assay for LNCaP cells after stimulation with androgen. The protocol I follow is I grow the cells in 96 well plate in 10% complete medium for 48 hrs. then remove the medium, give a PBS rinse of the plate and then add medium with 2% charcoal stripped serum. After adding medium I stimulate with different doses of androgen. However, the problem is, since LNCaP cells are loosely adherent cells, they tend to come out when I aspirate medium or rinse with PBS. They also come out when I add fresh medium with 2% CS FBS. As a result, the cell number varies.. which in turn will give me a wrong cell proliferation index. IF somebody cud help me on this topic..
Hi, Sheebs, have you solved your problem about lncap cells? I am doing experiments using lncap cells too, I faced the same problem as you, but I did it very carefully, the variation seems OK, but the effect of DHT is not so significant after 72hrs treatment(I use DHT as a postive control), how you did your experiment? what's your effect looks like? maybe we can exchange our opinions on lncap cells, what do you think?
Hi,, I grow MCF-7 in DMEM+10% FBS+ antibiotic and they are growing just fine.. they are kind of slow.. so what I do is increase the number of cells, i usually seed around 0.7 million cells in p-25 and they grow to full confulency within a week. you just control the number of cells seeded.
I never change the media the next day, usually do that the third day. and I never added insuline.. and they are growing just fine... and mine grow in epithelila layer form not doms
I also grow them in RPMI-1640 with 10% FBS, they grow well, the doubling time is silimar to ATCC reported. but when I set up the 96-well plate, treat with various doses of estradiol, they didn'grow well, and there are more and more dead cells,I have no idea what happened to these cells, Dose anyone have any idea? I repeated twice, same thing happened.Can anyone tell me how you do the experiment of MCF-7 cells treated with 17-beta estradiol?