Hello All,
I'm working on SHSY5Y cells differentiation. My problems are:
1-After adding the final differentiation media to the cell cultures, they started to detach from the plates, and became floating day by day. However, I've coated the plates previously by Poly-D-Lysin.
2- After adding the final differentiation media, cells look clumped.
Can someone suggest me differentiation protocol or any suggestion to solve my problems.
Thanks in advance.
Hi Elham Amini. E.A
1. Differentiate the cells in the flask itself if you are facing problems with detachment. It may be due to uneven coating or not washing.
2. SHSY5Y cells often get clumped and grow over them because of improper mixing during the passaging time.
*Grow the undifferentiated cells in 2% FBS media containing 10uM Retinoid acid. Change the media every 2 days. The cells should be differentiated in 10-12 days.
Thanks,
Samir Ranjan Panda
Hello Samir, Can we plate the cells in 96 well plates after differentiating in the flask by using trypsin?
Have you tried differentiation in the 60mm dishes?
Hi Harish
* I haven't tried differentiating the cells in the dish. Maybe you can give it a try if that works that would be best.
* And yeah we can plate the cells in 96 well plate however we need to plate in 2% FBS media containing RA.
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