2 replies to this topic

[Marvilla]

Hello,

I have a problem with LNCaP cells. After cell passaje they do not disgregate well and they form ugly clumps(see attached photo). I've tryed to increase trypsin concentration (2X), time of trypsisization (10min), volume of trypsin (2ml per 10cm plate) and even i've vortexed them but cell look like in the attached photo. I'm wondering if those clumps are normal? What do you think? Are the cells in the photo good to make and experiment?

Thanks.

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[bob1]

I have heard that LnCAP cells are quite difficult to culture properly.  Over trypsinising is one thing that can cause cells to clump, cell damage will also cause them to clump, so vortexing should never happen to cells.

[gilahari]

Marvilla, on 30 July 2010 - 06:50 AM, said:

Hello,

I have a problem with LNCaP cells. After cell passaje they do not disgregate well and they form ugly clumps(see attached photo). I've tryed to increase trypsin concentration (2X), time of trypsisization (10min), volume of trypsin (2ml per 10cm plate) and even i've vortexed them but cell look like in the attached photo. I'm wondering if those clumps are normal? What do you think? Are the cells in the photo good to make and experiment?

Thanks.

LNCaPs are very sensitive to media and serum conditions as well as how you culture them. The picture that you have attached does not look good. In my opinion, you should start over. The serum that has worked well for me is FB-02 from Omega Scientific. RPMI with 10% serum + P/S. Trypsinize for 3-4 minutes and split at 1:3 ratio. The cells should attach and flatten out nicely overnight. If not, then they are not doing well. Hope this helps.