Hi All;
I am growing PC12 cells and trying to do a secretion assay with Nicotine induction. I have done it couple of times already but not getting enough induction with nicotine. I have tried with different aliquots of cells and also used new Nicotine stock as well. Without nicotine induction I can not really go to the next step of my experiment, so I am kind of stuck. Any suggestion? Thanks in advance.
Saiful.
Adherent PC-12 Adh ATCC ® CRL-1721.1 works better than the normal floating PC-12 ATCC ® CRL-1721. You may try with adherent version.
Jia-da,
Thanks. I am using adherent ones only. Those are the ones stored in lab. Anything else?
Saiful.
may be your cell line modified after long passage (it commonly happens with carcinoma cell lines). it may contains very less nAChR (nicotic receptor). You may transfect your cell with nAChR Plasmid.
Hello, I want to order PC-12 Adh ATCC but I found the flask which has been recommended by ATCC is so expensive. how you guys came over that?
@ Saiul, What is your passage #? From my experience PC12 cells are tricky to handle. As Dr. Gayen stated, PC12 cells do change phenotypes as they get older. I recommend you the classic PC12 cells article by Dr. Greene & Dr. Tischler. It helped me to tweak a few parameters to gain more control over the cultures. I have been working with PC12 cells with a 95% of them in suspension and a few of them adhered weakly. I use PC12 cells to perform transfections and NGF inductions. If your current vial of cells do not respond, I would thaw a early passage and see how they respond.
Hopefully this information helps you. Good luck :)
http://www.ncbi.nlm.nih.gov/pubmed/1065897
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