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. Author manuscript; available in PMC: 2020 Jun 26.
Published in final edited form as: J Vis Exp. 2019 Oct 24;(152):10.3791/60346. doi: 10.3791/60346

Figure 1: Measuring organoid formation rates of human and mouse prostate cells.

Figure 1:

(A) Schematic overview of cell resuspension in basement membrane matrix (left) and organoid seeding in matrix domes (right). (B) Relative organoid formation of human (CD49f+)-derived basal and (CD26+)-derived luminal cells (%, y-axis; mean ± SD) in the presence of 1 nM DHT. A total of 200 cells were seeded and the number of organoids was quantified 7 days post-seeding (n = 3, ***p < 0.01, t-test). (C) Relative organoid formation of murine WT, PtenΔ/Δ, P53Δ/Δ, and P53Δ/Δ PtenΔ/Δ organoids (%, y-axis; mean ± SD) in the presence of 1 nM DHT. A total of 200 cells were seeded and the number of organoids was quantified 7 days post-seeding (n = 3). p53 and Pten loss was mediated by the gRNA’s targeting of the p53 and/or Pten locus in organoids expressing Cas9 constitutively under a Rosa26 promoter.