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
- - - - -

Feeder cultures


  • Please log in to reply
2 replies to this topic

#1 LostintheLab

LostintheLab

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 62 posts
2
Neutral

Posted 28 January 2009 - 05:01 PM

*Tears of joy and relief flowing down my face*

Its back!

Anyway,

I have some cells that I need to culture with a feeder layer- I have irradiated fibroblasts and keratinocyte cell line. Since the fibroblasts are irradiated I have to replace them every 2-3 weeks, but my keratinocytes should be confluent before then.
Do any of you think I can seed my fibroblasts in a few different flasks- to act as a stock for the keratinocyte cell line or will I have to seed fresh fibroblasts each time before spliting my keratinocytes?

Any advice- I've never used a feeder layer before and can't afford to buy these cells for a while so I don't want to mess things up.


Lost
I knew it! I knew it! Well, not in the sense of having the slightest idea, but I knew there was something I didn't know.

#2 gugo

gugo

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 30 January 2009 - 05:10 AM

I have work a little with feeder layers. I didn't irradiated the cells, I used mitomycin to inactivate their growth. I froze little aliquots of my fibroblast cell line and I only treated the cells when I need, after thaw them and seed.
I supose your cells irradiated didn't growth, but I don't know if you could mantain them in culture for long time.
If I were you I would buy a fibroblast cell line (I used 3T3-J2 from mouse) and then you work with them as usual and only irradiate o treat with mitomycin when necesary . After that, you add your keratinocytes over your feeder layer until confluence.
Best,
Gugo

#3 LostintheLab

LostintheLab

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 62 posts
2
Neutral

Posted 01 February 2009 - 04:35 PM

I have work a little with feeder layers. I didn't irradiated the cells, I used mitomycin to inactivate their growth. I froze little aliquots of my fibroblast cell line and I only treated the cells when I need, after thaw them and seed.
I supose your cells irradiated didn't growth, but I don't know if you could mantain them in culture for long time.
If I were you I would buy a fibroblast cell line (I used 3T3-J2 from mouse) and then you work with them as usual and only irradiate o treat with mitomycin when necesary . After that, you add your keratinocytes over your feeder layer until confluence.
Best,
Gugo



Hi thanks,
Unfortunately the cell line I got came with the already irradiated cells so I figured I should use them.
But I might buy the fibroblast cells and carryon that way.

Lost
I knew it! I knew it! Well, not in the sense of having the slightest idea, but I knew there was something I didn't know.




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

©1999-2013 Protocol Online, All rights reserved.