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. 2014 Jul 17;2(7):e12075. doi: 10.14814/phy2.12075

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© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Timeline of the differentiation protocol. (A) A step‐wise differentiation protocol was used to generate Foxa2+Nkx2.1+ endodermal lung progenitors at 20% and 5% oxygen cell culture conditions. DEM = definitive endoderm medium; ADEM = anterior definitive endoderm medium; LPM = lung progenitors medium; Nog = Noggin; SB = SB431542; WFKBE = Wnt3a, FGF10, FGF7, BMP4, EGF; F2/H = FGF2, heparin sodium salt. (B) On day 12, cell cultures were gently trypsinized and subcultured. On day 22, cells were analyzed for the expression of mature lung markers, such as pro‐surfactant protein C (proSPC).