transfection of zebrafish fibroblast cell lines?

[Shawn Burgess]

Jill:

We have had reasonable luck making stable lines in Pac2 cells 
(derived from dissociated 1K cell embryos). We transfect with 
Lipofectamine 2000 from Invitrogen. Although advertised that you 
can leave the lipofection reagents on the cells overnight, our 
experience is that it will kill 90% of the cells if you do that. 8h 
seems sufficient. I think we see around 10-20% of the cells are 
transfected (no hard data on that).  


Shawn




in article a5dof4$nlh$1 at mercury.hgmp.mrc.ac.uk, Jill at
jmurtha at umich.edu wrote on 2/25/02 11:24 AM:

> Hello!  I was wondering if anyone has successfully transfected any
> of the zebrafish fibroblast cell lines that are available from ATCC,
> or any other types of zebrafish cell cultures for that matter?
> Someone told me that fibroblasts are very difficult to transfect -
> is that true?  I haven't been able to find any references on
> transfection of any of the 4 fibroblast lines available at ATCC, but
> I thought I'd ask here.  Thanks in advance for any info.
> 
> Jill
> 

---
Shawn Burgess
Developmental Genomics Section
Genome Technology Branch
National Human Genome Research Institute
Bldg 50
50 South Dr.
Rm. 5537, MSC 8004
National Institutes of Health
Bethesda, MD  20892

ph: 301-594-8224
Fax: 301-496-0474
burgess at nhgri.nih.gov

http://www.nhgri.nih.gov/Intramural_research/People/burgess.html