Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Problem with growing HEK293

cell culture

  • Please log in to reply
2 replies to this topic

#1 erimyes

erimyes

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 09 April 2017 - 08:17 AM

I have noticed my cells are growing slower than regular a week ago and I suspect about contamination.So I did staining & imaging yesterday using hoescht 33342 dye and noticed abnormal shape of nucleus in my cells. Does anyone have any idea what could be causing this? I prepared my medium on 22nd of February with following recipe. Plus I was doing transient transfection to my cells previously gfp fused plasmid that I believed to be expressing, however I also noticed yesterday that my cells without any transfection are emitting flourescence under gfp  spectrum too. I have no prior knowledge working with cell cultures so any insight would be appreciated. TIA

Attached Thumbnails

  • 4_DAPI.png


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,343 posts
509
Excellent

Posted 10 April 2017 - 07:51 AM

The cells look fairly normal - nuclei are generally not round in adherent cells.

 

Growth slowing is another matter - if you can tell us what you have done with the cells and anything that might have changed (e.g. different lot of FBS recently), we will have a better idea of what might be going on. 

 

There is always some auto-fluorescence with cells in the green channel - and it is can be a big problem as it is almost impossible to remove. If you are looking for a faint change or minor differences in localization it might be better to go for a RFP or YFP. 



#3 Missle

Missle

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 166 posts
27
Excellent

Posted 10 April 2017 - 09:43 AM

I normally don't use media for much more than a week.  If you're still using complete media from 6 weeks ago, that's not a good thing.  What seeding density are you passaging them and what is the media formulation?







Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.