I seeded a flask of human neuroblastoma cells almost three weeks ago, and they did not grow to confluency yet. This is the second passage; the first time the cells took like 2 weeks to reach confluency. I split 1:4 three weeks ago, and i am still waiting.
Today i looked at the flask under the microscope and i saw that there are some very well-spread cells in some places, so i believe they are still alive. What can i do to help them grow more? The medium was fresh but i could prepare other medium. The medium was the same i used the previous time. Thank you for any imput!
Hi Sara,
I suggest you look at these aspects:
- Are you using proper medium for growing this cell line (DMEM/F12 1:1)? Is it within the expiration date?
- Is the culture medium at the proper pH?
- Are you using the appropriate concentration of fetal bovine serum? (10% FBS)
- Is the incubator working properly (37°C and 5% CO2)?
If all observed aspects are being followed, I suggest thawing a new vial.
https://www.atcc.org/products/all/CRL-2266.aspx#culturemethod
you can verify in this website
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