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Cell proliferation assay for LNCaP cells


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#1 sheebs

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Posted 06 January 2010 - 11:42 PM

Hi,

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..

Edited by sheebs, 07 January 2010 - 12:41 AM.


#2 bob1

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Posted 07 January 2010 - 03:22 PM

try coating the plate with a cell attachment factor such as collagen (gelatin) or fibronectin.

#3 shuiyi

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Posted 09 January 2010 - 09:29 PM

i usually use polylysine to precoat the plate, and this maybe help some.

http://www.genlantis..... Protocol.pdf

Edited by shuiyi, 09 January 2010 - 09:32 PM.


#4 isabellaweng

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Posted 29 August 2010 - 05:21 PM

Hi,

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?

#5 liezl

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Posted 08 May 2013 - 05:25 AM

years later and I have the same questions...did you succeed? :)




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