Can EDTA remove fibroblast cell? - (Nov/12/2006 )
Hello,all
when I do epithelial cell primary culture,however,fibroblast cells overgrow.I saw someone suggest before subture with trypsin,wash the cells twice with EDTA. Dose anyone konw can EDTA remove fibrolbast or leave the fibroblast still in the plasticware?When subculturing,I use 0.25%trypsin+0.02%EDTA，but obtain few epithelial cell.
-wing111-
QUOTE (wing111 @ Nov 12 2006, 08:21 PM)
Hello,all
when I do epithelial cell primary culture,however,fibroblast cells overgrow.I saw someone suggest before subture with trypsin,wash the cells twice with EDTA. Dose anyone konw can EDTA remove fibrolbast or leave the fibroblast still in the plasticware?When subculturing,I use 0.25%trypsin+0.02%EDTA，but obtain few epithelial cell.
when I do epithelial cell primary culture,however,fibroblast cells overgrow.I saw someone suggest before subture with trypsin,wash the cells twice with EDTA. Dose anyone konw can EDTA remove fibrolbast or leave the fibroblast still in the plasticware?When subculturing,I use 0.25%trypsin+0.02%EDTA，but obtain few epithelial cell.
Hi, I have relatively less experience in this field; but I do know that EDTA is used as a agent for cell dissociation when working with very delicate cells that are very susceptible to trypsin and are not too adherent. Probably since fibroblasts are more adherent EDTA would not detach them soon. Try a 2mM solution of EDTA; incubate your plaet/flask for an appropriate length of time and check the cell population. If thats too weak try increasing to 5mM EDTA. I hope this helps.
-smartsunny-