Glycyrrhizin (GL) and cell culture - (Sep/07/2005 )
I was trying to setup some cell assay to see the possivle effect of GL on activating/inactivating various cell lines (e.g. MCF-7, RAW264.7, RBL-2H3,...etc.). Am wondering if anyone familiar with this chemical could offer some suggestion on the problems I had encountered.
First, I tried to dissolve GL in either dH2O or PBS and found it simply didn't go into these two reagents at all. Admittedly, I should have consulted with its MSDS first.. So, I finally was able to dissolve it in either acetic acid (pH2.4) or ammonia (pH11.6) at 2mg/ml and 100 mg/ml, respectively.
However, here comes the delimma: when I added GL-acetic acid or GL-ammonia into my cell cultures (at final of 100x to 10x dilution from the stocks; cultures were 200ul/well in a 96-well plate), I found cells were obviously NOT happy with either acetic acid or ammonia present even at the lowest 10x dilution. With the GL-ammonia added cells, the media remains its normal color (indicating no significant pH change) and the cells were all shrunk; while with the GL-acetic acid added cells, the media turned into yellowish color right off and for adherent cells, they all look round (not the normal spindle adherent looking). Obviously I couldn't do any experiment with GL-ammonia preparation, nor did I have enough confidence on GL-acetic acid preparation since the cellular morphology seemed changed with the presence of acetic acid.
I've been looking up the Materials & Methods sections on numerous research articles (GL has been done on numerous anti-viral/anti-tumor studies); however, they didn't seem to mention the detailed preparation of GL and its use (e.g. dilution) in their cell line studies. I would gratefully appreciate if any expert could help me out with this obstacle.. Many thanks in advance!!
Generally I make a stock of 1000x assay conc. sothat it gives enough dilution of any solvent used. You could probably try this out.
Another thing I would like to suggest is to approach the authors who have worked in this field (you should be able to do this from their research papers).
Hope it helps.