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
- - - - -

Salamander Mueller Glia


  • Please log in to reply
No replies to this topic

#1 Kung_UMDNJ

Kung_UMDNJ

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 28 June 2010 - 07:22 AM

Hi all,

I need to grow and maintain Salamander(Ambystoma tigrinum) Mueller cells for a week. I was wondering if anyone had experience culturing amphibian Glial cells and if they had any protocols or tips. Currently I can get a relatively (~60-70%) pure isolation, but 99% of them die within a 3 days. I currently grow them in Salamander Media: 108 mM NaCl, 2.5 mM KCl, 2 mM HEPES, 1 mM NaHCO3, 1.8 mM CaCl2, 0.5 mM NaH2PO4, 1 mM NaHCO3, 24 mM glucose, 0.5 mM MgCl2, 1 mM Na pyruvate, 7% medium 199, 1x minimum essential (MEM) vitamin mix, 0.1x MEM essential amino acids, 0.1x MEM nonessential amino acids, 2 mM glutamine, 2 g/mL bovine insulin, 1 g/mL transferrin, 5 mM taurine, 0.8 g/mL thyroxin, 10 g/mL gentamicin, and 1 mg/mL bovine serum albumin (pH 7.7). Cultures are maintained at 10C. I have also tried: increasing K+ to 15 mM, decreasing glucose to 0.5 mM, adding 10% FBS, changing O2 levels by increasing and decreasing amount of culture media.
Thanks for any help.




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

©1999-2013 Protocol Online, All rights reserved.