Dr. Haron,

I believe all the information you need is in Encinas et al., J. Neurochem. 75:991, 2000. If you want to keep your cells alive, as per this report, simply maintain them in serum-free medium and in the presence of the neurotrophin--PC12 and NS-1 cells behave similarly after differentiation with NGF. I would suggest to you also the often-underrated expedient of contacting these authors directly for further information. Thanks for the opportunity to consider this interesting question. You might also consider that maintenance of human neuronal cells via an iPSC avenue, rather than differentiation of a human tumor cell line, might be more intellectually rewarding and translationally impactful, unless of course your focus is neuroblastoma neurobiology per se.

Best regards,

L.E.E.

Thanks for the responses Lee and Shelanah

@Lee: I've re-read the paper that you've quoted - thanks for pointing the way. I'll discuss with my fellow colleagues regarding the utilisation of neurons differentiated from iPSCs. 

@Shelanah: I was planning to maintain them for 2-3 weeks whilst I synthesize my materials to test on them. After reading the paper suggested by Lee, I'll use BDNF in serum-free media to maintain them. 

Another question: Is it advisable to (i) trypsinise and seed these diff-Sy5y from flasks/dishes into 96-well plates, or would it be better to (ii) differentiate them directly in the plates? One issue I have with (ii) is that the cell density of the diff-Sy5y after a whole week of RA and BDNF treatment might not be the same in each well, correct? At least with (i) I can control the number of cell I seed in each well to ensure that whatever assay I run on them would yield a more homogenous result.

Thanks.

Muhammad

hi Muhammad, 

I am working on SH-SY5Y cells. 

 i agree with you that the numbers of cells after differentiation in each well are not the same. 

recently, i differentiate cells in dish then trypsinised them and seed. however, differentiated SH-SY5Y cells are not easy to trypsinised comparing to the undifferentiated one. therefore, i incubated the cell 2min more.

what is your approach ?