I have a few cell clones of NIH-H23 engineered to express SpCas9. These cells grow very slowly compared to other cancer cell lines (e.g. A549) and following thaw to expand clones they detach from plates readily and grow only in low density. I am planning on testing PDL-coating and Gelatin for 2D culture to establish a kill curve for puromycin, are there any established culture conditions that I am overlooking? I plan to culture 3D by adding methylcellulose to the media and reduced-matrigel coating.

2D/Base Media is RPMI-1640 w/Glutamax + 10%FBS + Pen/Strept.

I have not ruled out potential mycoplasma contamination yet, I am testing shortly.

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