(q)HepG2 care and feeding

David L. Haviland, Ph.D. HAVILAND at KIDS.WUSTL.EDU
Fri Jun 16 12:22:12 EST 1995


In <jbdaniel.46.0015E9C8 at facstaff.wisc.edu> jbdaniel at facstaff.wisc.edu writes:

> Hi-
> 
> I am wondering what HepG2 cells require for routine growth.  Is DMEM/ 10% 
> fetal calf supplimented with anything?  (pyruvate, glutamine, bicarb, 
> nonessen. amino acids, insulin, etc).  Is high glucose recommended for the 
> DMEM?  Passage with trypsin/ EDTA?  
> Any input would be appriciated.  
> Josh


Josh:

HepG2 cells will grow relatively well in just DMEM with 10%FCS, 
supplemented with extra l-glut.  Traditionally, I'll "sweeten" that with 
pyruvate and NEAA.  We do use the high glucose DMEM as well (hybridomas, in 
my hands, seem to do better in that than standard RPMI1640).  Yes, pass 
them using trypsin/EDTA.  HepG2's go "forever" in culture but for us have 
often gone through phenotypic changes so make sure every now and then that 
your stock of Heps continue to do what you want.

Personally, I feel as though Heps do a bit better in MEM-alpha with 10%FCS, 
glut, and pyruvate (antibiots if you prefer), than in DMEM.  No, I 
*haven't* done any growth curves but after working with them for the last 5 
years, I think they grow a bit better in MEM-a than DMEM but DMEM will do 
just fine.

Hope this helps,
David





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