I'm going to use the RBL-2H3 cell line for my in vitro study involving the inhibitory activity of honey bee product on mast cell degranulation. I just want to know if there is anyone who has experience dealing with this type of cell? Is it easy/difficult to handle? I would be glad if someone can share their precious experiences working with this cell.
Dear Dr. Suhana,
Yes, I worked with RBL2H3 cells for the last 20 years.
It is a rat basophilic leukemia-cell derived cell line. The cells carry many beneficial characters as a mast cell, especially responding to mast-cell secretagogues. Instead, I used the cell line as a serotonin-producing cell line. As to the cell handling, RBL2H3 cells are somewhat unique but manageable. What do you want to know with the cell line? Take a look some of my publications and make narrower your question.
Hiro
1) Hasegawa et al (1996) Stimulation of tryptophan hydroxylase production in a serotonin-producing cell line (RBL2H3) by intracellular calcium mobilizing reagents. FEBS Lett 392, 289-292.
2) Hasegawa et al (1999) Iron dependence of tryptophan hydroxylase activity in RBL2H3 cells and its manipulation by chelators. Eur J Biochem 261, 734-739.
3) Iida et al (2002) Proteasome-driven turnover of tryptophan hydroxylase is triggered by phosphorylation in RBL2H3 cells, a serotonin producing mast cell line. Eur J Biochem 269, 4780-4788.
4) Sawabe et al (2008) Cellular uptake of sepiapterin and push-pull accumulation of tetrahydrobiopterin. Mol Genet Metab 94, 410-416.
Dear Salme,
I have studied the inhibition effect of various honeys on the release of beta-hexoaminidase from IgE- sensitised and non- sensitised RBL cells. The data were presented at EMBRN meeting 2011 in UK. We got our RBL cells and details of culture methology from Dr L Vogel and Prof Vieths in Germany.
Dear all,
Thank you for your kind reply. Actually i'm interested in studying the inhibitory effect of my sample against the mediator release of mast cell such as histamine and beta-hexosaminidase. I will use Elisa kit for histamine and 4-nitrophenyl N-acetyl- β-D-glucosaminide for beta hexosaminidase assay. The antigen will be DNP-HSA. Based on your experience, is the result for this type of mediators reproducible? Since no one in my lab had experience dealing with this type of cell so if possible I would like to know about the passage and the confluency so that I can get a rough idea about the cell.
Histamine assay will be expensived unless you got histamine analyser. beta- hexoamindase assay are cheaper and very reproducable if you don't use too many cells in the wells.
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