Hello everyone,
In my upcoming project, I will use poly-L-lysine coated culture flasks due to the culture conditions of neural stem cells.
One thing that comes to my mind is if I need to perform a viability test such as MTT or SRB on these cells, do the 96-well plates have to be poly-L-lysine coated as well? Buying manufactured coated plates increases the cost a lot. I am guessing my deviations may increase if I coat them myself. Also, does poly-L-lysine interfere with these kinds of tests? I do not have much experience with this subject. I would be very happy if you share your information not only for the viability tests but also for any kind of plate-based tests for the neural stem cells.
Thank you very much.
Dear Gülşah Torkay ,
For your first question, I think yes you should coat the 96-well plates with poly-L-lysine as well because in MTT assay, for example, you will seed your cells overnight so that they can attach to the plates and grow. This poly-L-lysine will help the neural stem cells to attach better to the flask and will increase their adhesions because neural stem cells don't attach to plates or flasks easily.
For your second question, as far as I know, coating the plates with poly-L-lysine will not interfere with cell viability tests, there are some articles that coated plates with poly-L-lysine and did the MTT or other viability tests.
Also, in order to reduce the risk of your deviation when coating the plates with poly-L-lysine, I recommend following the protocol provided by Sigma :
1. Add 10 ml sterile tissue culture grade water to 1 mg of poly-L-lysine(CAT:7280),
NOTE: (this step is not necessary for poly-L-lysine Solutions(like cat: P4707))
2. Coat surface with 1ml/25 cm2
3. Rock gently
4. After 5 minutes, remove solution and rinse with sterile culture grade water
5. Allow drying at least 2 hours before using.
Hope this can be helpful for your task.
Dear Gülşah Torkay ,
You're welcome and good luck with your project!
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