I'm working with the cell line CRL-1446, which should beat in culture with the addition of acetylcholine (according to ATCC). We culture in 1% FBS DMEM to form myotubules as suggested by ATCC. We've tried adding calcium, norepinephrine, and acetylcholine with no luck.
1% FBS seems low, would you consider 10%? What conc of Ca?
Why both norepinephrine and acetylcholine, seems like they would have contradicting effects that could raise and decrease rate respectively. However, they should be beating spontaneously first.
Hello,
I agree with Rafael that 1 % FBS is low, ATCC actually sugests 10 % FBS in the medium. This is what ATCC says regarding beating of the cell line you use (I copied it from their webpage):
H9c2(2-1) cells (ATCC® CRL-1446) will fuse to form myotubes and will respond to acetylcholine stimulation to beat. The ATCC tests each lot for the formation of myotubes. We do not test for acetylcholine stimulation, however other customers have tested for this property and reported the results to the ATCC. The ATCC does not have any information about whether they will beat spontaneously. The ATCC has not tested any of the other cardiac myocyte cultures in the collection for the ability to beat in culture.
Good luck!
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