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. 2020 Oct 21;27(6):890–904.e8. doi: 10.1016/j.stem.2020.10.005

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© 2020 Elsevier Inc.

Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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Establishment of Chemically Defined Human Lung Alveolosphere Culture System

(A) Schematic representation of human alveolosphere cultures and passaging in SFFF medium.

(B) Representative images of human alveolospheres from different passages. Scale bar: 100 μm.

(C) Quantification of the colony formation efficiency (CFE) of human alveolospheres at different passages.

(D) Immunostaining for SFTPC (green), SFTPB (red), and AGER (gray) (left panel) or SFTPB (green), HTII-280 (red), and DC-LAMP (gray) (right panel) at P1 and P3 human alveolospheres cultured in SFFF medium for 14 days.

(E) Immunostaining for SFTPC (green) and HTII-280 (red) in cells dissociated from alveolospheres at P2 (top) and P8 (bottom).

(F) Quantification of HTII-280⁺ SFTPC⁺ cells/total 4′,6-diamidino-2-phenylindole (DAPI)⁺ cells derived from alveolospheres dissociation from P2 (orange) and P8 (blue).

(G) Schematic representation of human AT2 to AT1 differentiation in alveolospheres. AT2s were cultured in SFFF medium for 10 days, followed by culture in ADM for 14 days.

(H) Immunostaining for SFTPC (green) and AGER (red) in human alveolospheres cultured under ADM condition for 14 days. Scale bars: 100 μm (B); 50 μm (D); 20 μm (E); 20 μm (H). DAPI (blue) shows nuclei in (D), (E), and (H). Data are presented as mean ± SEM.