Hi all
We imported a tube of hybridoma cell line, but the courier company accidentally forgot to top up dry ice, while my cells were kept at -10 degree for approx 2 days. I tried to put the cells back into culture as soon as I received them, and approximately 10% of the cells survived.
As the supplier suggested, the cell should be maintained with RPMI1640-10%FBS. I tried to bulk them up again, but they seem to remain dormant (viable but barely proliferate).
I have tried to add some b-mercaptoethanol to promote cell proliferation but seems not very helpful. Does anyone have some suggestions?
It is very difficult to get the import permit and very expensive to ship the cell . Therefore, I would prefer not to ship the cell again.
Dear Dr. Gaoqian Feng
I have a few suggestions for you.
Since you have just 10% of the cells left, culture them in a culture vessel having a small surface area so that the cells are in close proximity with one another. Secondly, supplement the growth media (RPMI-1640 +10%FBS) with 4mM L-glutamine, 0.1mM MEM vitamins, 0.1mM non-essential amino acids and 1mM sodium pyruvate.
Supplementing the growth media with amino acid solution will help increase the cell density.
Regards,
Malcolm Nobre
Rescuing a hybridoma cell line involves reviving cells from frozen stocks and culturing them to re-establish a viable cell population. Here's a step-by-step guide on how to rescue a hybridoma cell line:
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