Hello, I'm trying to culture my N2a cell line but I have proliferation problem. Their proliferations are so slow. I started my culture 5 days ago. I use EMEM, %10 FBS and penicillin-streptomycin (because of company's advices). How many passages do I need for proliferation? I also wondered whether they contaminated or not. Because I saw different shaped things and I'm sharing them. Do I need anything else for proliferation? Also I don't want to differentiate them I just want to proliferate. Can someone help me?
First pic: 3rd day
Second pic: 4th day
3-4-5-6. pic: 5th day of my cultures
Thanks,
Arda.
I think the seeding density is low. For some cells, if its lower than a threshold, they do not grow.
Hello Arda,
Do you grow the cells in flasks.If so try growing in T-150 flask.Allow them to settle for about 6-7 hrs (give enough time for them to attach to the flask), then change the media.
I have tried this for HT 1080 cell line.I had the same problem with their proliferation when I was growing them in T-75.Then I used T-150 and changed the media after 6 hrs of placing the cells in the flask.They were confluent enough within 3-4 days compared to 8-9 days when using T-75.
Hope this helps.
All the best.
I think you should inreases amount of l-glutamin and may be amount of fbs... you can open a new stok and after 24 hour you can change medium... also you can prepare a new EMEM. I think you can test your cells both t-25 and t-75 for first step. ıf your sample is good, you can pass pasage 3 or 4 day. but your sample is not good, you can wait several days and only change dmem (slowly). hope this helps... good times, good work
I think the seeding density is low. For some cells, if its lower than a threshold, they do not grow.
I am now having the same problem. Were you able to increase the proliferation? What worked for you?
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