Skip to main content
. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Science. 2020 May 1;368(6490):497–505. doi: 10.1126/science.aay0267

Fig. 3. Enhanced regenerative potential of murine luminal cells after castration in organoid culture and in vivo.

Fig. 3.

(A-C) Enhanced organoid formation of L1 and L2 cells isolated post castration or during regeneration. (A) Relative organoid formation (%, y axis; mean ± standard deviation) in the presence of 1 nM DHT in cultures initiated by L1 or L2 cells isolated by CD26/Dpp4 (L1), CD133/Prom1 (L1) or Sca1/Ly6a (L2) expression from hormonally intact prostate (blue) or prostate 28 days post castration (red). The number of organoids was quantified 7 days after seeding 200 cells. (N=3 * designates p< 0.05, t-test). (B) Relative organoid formation (%, y axis; mean ± standard deviation) from L1 or L2 cells isolated by CD26/Dpp4 or Sca1/Ly6a expression, respectively, from a prostate 28 days post castration (red) or a prostate 2 days into regeneration (blue) in the presence or absence of 1 nM DHT. The number of organoids was quantified 7 days after seeding 200 cells. (N=3 * designates p< 0.05, t-test). (C) Representative images of organoids derived from CD26/Dpp4+ L1 cells (top) or Sca1 L2 cells (bottom). Left: brightfield; Right: Confocal images (single Z and maximum projection) stained with Ck8 (red), Ck5 (green), Epcam (white) and DAPI (purple) 7 days post establishment. Scale bars: 100μm. (D) Lineage tracing strategy. (E, F) Contribution of multiple clones to prostate gland regeneration. (E) Top: Maximum projection of a castrated prostate 7 days post tamoxifen injection (left) and 4 weeks post regeneration (right). Only red fluorescence protein (RFP) and yellow fluorescence protein (YFP) are shown. Scale bars: 500μm. Bottom: Higher magnification of lineage-traced prostates showing contribution of multiple clones to gland regeneration. Scale bars: 100μm. (F) Distribution of size of different color clones. Log2 clone size (y axis) is plotted from three independent mice compared to control (castrate 7 days post tamoxifen) (x axis). Raw data is in table S2. As observed previously, GFP+ clones are infrequent in the prostate (34).