biostudies-literature00580Jacob ABLRD VANCATS NIH HHSCystic Fibrosis FoundationNIEHS NIH HHSNHLBI NIH HHSCReMNIHNIGMS NIH HHS472-488.e10https://www.ebi.ac.uk/biostudies/studies/S-EPMC5755620biostudies-literatureUnknown21(4)Lung alveoli, which are unique to air-breathing organisms, have been challenging to generate from pluripotent stem cells (PSCs) in part because there are limited model systems available to provide the necessary developmental roadmaps for in vitro differentiation. Here we report the generation of alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of lung alveoli, from human PSCs. Using multicolored fluorescent reporter lines, we track and purify human SFTPC+ alveolar progenitors as they emerge from endodermal precursors in response to stimulation of Wnt and FGF signaling. Purified PSC-derived SFTPC+ cells form monolayered epithelial "alveolospheres" in 3D cultures without the need for mesenchymal support, exhibit self-renewal capacity, and display additional AEC2 functional capacities. Footprint-free CRISPR-based gene correction of PSCs derived from patients carrying a homozygous surfactant mutation (SFTPB^121ins2) restores surfactant processing in AEC2s. Thus, PSC-derived AEC2s provide a platform for disease modeling and future functional regeneration of the distal lung.Cell stem cellDifferentiation of Human Pluripotent Stem Cells into Functional Lung Alveolar Epithelial Cells.PMC5755620R01 HL095993R24HL123828U01HL134745R01 HL111449P30 ES013508U01HL099997U01TR001810R01HL128172R01 HL136722R01 GM108807TL1TR001410F31 HL134274HL059959F31HL134274TL1 TR001410DAVIS15XX1I01 BX001176T32 HL007035U01 HL134745R01 HL122442U01 HL099997R33 HL120760U01 HL134766R01 GM120060HAWKIN15XX0UL1 TR001430R01 HL132999R01HL095993F31HL129777R01 HL128172K08 HL130582U01 TR001810R01 HL119436R56 HL059959R24 HL123828F31 HL129777DAVISGO1UL1TR001430R01 HL059959R01HL122442R33HL120760U01HL134766Beers MFGuttentag SHCole FSDavis BRWambach JWhite FVHeins HTraber KKook SQuinton LJJacob AOchs MCrane AJean JCNa CLMorley MKotton DNMcCauley KBHawkins FWhitsett JAHurley KVedaie MZacharias WMorrisey EEWeaver TERusso SJHinds A58falseDifferentiation of Human Pluripotent Stem Cells into Functional Lung Alveolar Epithelial Cells.Lung alveoli, which are unique to air-breathing organisms, have been challenging to generate from pluripotent stem cells (PSCs) in part because there are limited model systems available to provide the necessary developmental roadmaps for in vitro differentiation. Here we report the generation of alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of lung alveoli, from human PSCs. Using multicolored fluorescent reporter lines, we track and purify human SFTPC+ alveolar progenitors as they emerge from endodermal precursors in response to stimulation of Wnt and FGF signaling. Purified PSC-derived SFTPC+ cells form monolayered epithelial "alveolospheres" in 3D cultures without the need for mesenchymal support, exhibit self-renewal capacity, and display additional AEC2 functional capacities. Footprint-free CRISPR-based gene correction of PSCs derived from patients carrying a homozygous surfactant mutation (SFTPB^121ins2) restores surfactant processing in AEC2s. Thus, PSC-derived AEC2s provide a platform for disease modeling and future functional regeneration of the distal lung.2017-01-01T00:00:00Z2017 Oct2024-10-16T13:42:16.303Z2019-03-26T23:58:04ZS-EPMC57556202896576610.1016/j.stem.2017.08.014