Hi everyone,
I've been working on differentiating human iPSCs to derive a pure and uniform culture of neural progenitor cells (NPCs). However, I'm encountering a significant issue during the neural rosette selection phase. Despite using the Neural Rosette Selection Medium (from Stem Cell Technologies) and manually picking the rosettes, I still end up with non-NPC cells contaminating the culture. This heterogeneity is adversely affecting the subsequent neuronal differentiation process.
Has anyone else faced this problem? What strategies or techniques did you find effective in ensuring a more pure NPC culture? Any tips on improving the rosette selection process or alternative methods to minimize non-NPC contamination would be greatly appreciated.
Thank you.
Change to forward programming with NGN2 overexpression either with gene targeting or piggybac strategies (constructs are available in Addgene) and forget about rosette pick up, I suggest.
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