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. Author manuscript; available in PMC: 2018 Dec 7.
Published in final edited form as: Cell Rep. 2018 Oct 16;25(3):598–610.e5. doi: 10.1016/j.celrep.2018.09.072

Figure 1. TGF-β Signaling Inhibition, ROCK Inhibition, and Low Calcium Synergistically Support Long-Term Epithelial Cell Proliferation.

Figure 1.

(A) Small molecules inhibiting the TGF-β signaling or ROCK supported the proliferation of late-passage PrECs/nRFP cells in the absence of feeder cells in the F medium. Data are represented as mean ±SD, n = 4.

(B) Synergy between A83-01 and Y-27632 in the F medium (four replicates per condition).

(C and F) PrECs/nRFP cells proliferated for 10 PDs in the F medium plus Y-27632 and A83-01 (F+Y+A) but continued to proliferate in the CR condition (C). Many cells in F+Y+A exhibited differentiated morphology (F).

(D) ROCK inhibitors synergistically promoted the proliferation of HBECs/nRFP cells in KSFM plus 1 μM A83-01.

(E) Synergy between A83-01 and Y-27632 in KSFM (four replicates per condition).

(G) Morphology of HFKs over successive passages in KSFM (P1 and P5) or the EpiX medium (P2, P11, and P20).

(H) TP63 was ubiquitously expressed in late-passage HFKs (P16) cultured in the EpiX medium.

(I-L) Expansion of HFKs (I), PrECs (J), HBECs (K), and mammary epithelial cells (L) in KSFM or EpiX.