I never handle the SHSY5Y cell line before, so it is a new thing for me, as far as I observed for these past five days, their growth is really slow. Is it normal? I use the complete media of nutrient mixture F12 Ham supplemented with 15% FBS (recommendation from ECACC), I'm afraid if there might be something I missed or if there is any recommended suggestion for me. Thank you.
They are very slow growing with a doubling time of ~70 hours, which is very much slower than most tumor cell lines I know.
Hello Dasilva Nuswantari
What seeding density have you used?
You should seed 2x10^6 cells per T-75cm^2 flask. The doubling time of SHSY5Y cell line is around 48-50 hours. If you seed around 2x10^6 cells/ T-75cm^2 flask, you should achieve 80-90% confluency in around 5-6 days. I had used Ham's F12: EMEM (1:1) supplemented with 15% FBS. I also added supplements such as L-glutamine (2mM) and 1% NEAA to the culture media. Why don’t you try this out?
Best.
nice information! thank you.. Philip G Penketh Malcolm Nobre
well, the number of my seeding density is 4x lower than your recommendation, that's why it took so long for them to reach confluency (a week-long w/ approximately only 65-70% confluency). I also use the exact same growth media as you suggested anyway..
is it the most suitable media for their growth? have you tried to grow them in Complete DMEM? I'm just wondering if maybe they have an alternative growth media that also suitable for supporting their growth Malcolm Nobre
Dasilva Nuswantari no, I haven't tried growing them in complete DMEM. You may try DMEM, but increase the seeding density.
Hi Dasilva Nuswantari
There are some precautions you need to take while handling SHSY5Y cells due to its slow growth properties.
* Please grow them in 15-20% FBS.
* Always grow at high seeding density, as these cells require exchange of nutrients and cell to cell connection to enable its growth.
* You can grow them in F12 media and supplement with Sod Pyruvate and Glutamax.
* Before experimentation, it is recommended to differentiate in 10uM RA in reduced serum media (2% FBS).
Thanks,
alright, thanks a lot for the nice recommendation!! I will keep in mind for future experiment Samir Ranjan Panda Malcolm Nobre
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