1ce4f2819b776488ec89c88f4a125ee8 pmc_9100165.pdf bb163af1fb86f684ec3fb2cf3af9558b3c75073b pmc_9100165.pdf a8ba0309af06e6318b9990d8288d4d64e6eb551e1f1ed89f0767d256cb1794de pmc_9100165.pdf Title: Lung Extracellular Matrix Hydrogels Enhance Preservation of Type II Phenotype in Primary Alveolar Epithelial Cells Subject: One of the main limitations of in vitro studies on lung diseases is the difficulty of maintaining the type II phenotype of alveolar epithelial cells in culture. This fact has previously been related to the translocation of the mechanosensing Yes-associated protein (YAP) to the nuclei and Rho signaling pathway. In this work, we aimed to culture and subculture primary alveolar type II cells on extracellular matrix lung-derived hydrogels to assess their suitability for phenotype maintenance. Cells cultured on lung hydrogels formed monolayers and maintained type II phenotype for a longer time as compared with those conventionally cultured. Interestingly, cells successfully grew when they were subsequently cultured on a dish. Moreover, cells cultured on a plate showed the active form of the YAP protein and the formation of stress fibers and focal adhesions. The results of chemically inhibiting the Rho pathway strongly suggest that this is one of the mechanisms by which the hydrogel promotes type II phenotype maintenance. These results regarding protein expression strongly suggest that the chemical and biophysical properties of the hydrogel have a considerable impact on the transition from ATII to ATI phenotypes. In conclusion, culturing primary alveolar epithelial cells on lung ECM-derived hydrogels may facilitate the prolonged culturing of these cells, and thus help in the research on lung diseases. Keywords: extracellular matrix; hydrogels; alveolar cells; type II phenotype; YAP Author: Esther Marhuenda, Alvaro Villarino, Maria Leonor Narciso, Marta Camprubí-Rimblas, Ramon Farré, Núria Gavara, Antonio Artigas, Isaac Almendros and Jorge Otero Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Apr 28 12:25:13 2022 CEST ModDate: Thu Apr 28 12:28:16 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4625190 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NBDQEL+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 KORLVE+VnURWPalladioL Type 1 Custom yes yes no 15 0 XGXOVH+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 VUVSDN+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 FKAKIX+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 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edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Children: Item: Title: Primary Alveolar Epithelial Cells Form Monolayers on Lung-Derived Hydrogels Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Culture of Alveolar Epithelial Cells on Lung-Derived Hydrogels Preserves the Expression of Type II Markers for Longer Periods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Culture of Primary Alveolar Epithelial Cells in Lung-Derived Hydrogels Inhibits Type II-to-Type I Transdifferentiation by Altering the Hippo/Rho Pathway Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Primary Alveolar Epithelial Cells Cultured on Lung-Derived Hydrogels Can Be Subcultured Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Decellularized Lung Extracellular Matrix Hydrogels Preparation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Primary Alveolar Epithelial Cells Isolation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Rho Kinase Inhibition Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Reverse Transcription qPCR Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Immunohistochemistry and Image Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Info: Title: Lung Extracellular Matrix Hydrogels Enhance Preservation of Type II Phenotype in Primary Alveolar Epithelial Cells Author: Esther Marhuenda, Alvaro Villarino, Maria Leonor Narciso, Marta Camprubí-Rimblas, Ramon Farré, Núria Gavara, Antonio Artigas, Isaac Almendros and Jorge Otero Subject: One of the main limitations of in vitro studies on lung diseases is the difficulty of maintaining the type II phenotype of alveolar epithelial cells in culture. This fact has previously been related to the translocation of the mechanosensing Yes-associated protein (YAP) to the nuclei and Rho signaling pathway. In this work, we aimed to culture and subculture primary alveolar type II cells on extracellular matrix lung-derived hydrogels to assess their suitability for phenotype maintenance. Cells cultured on lung hydrogels formed monolayers and maintained type II phenotype for a longer time as compared with those conventionally cultured. Interestingly, cells successfully grew when they were subsequently cultured on a dish. Moreover, cells cultured on a plate showed the active form of the YAP protein and the formation of stress fibers and focal adhesions. The results of chemically inhibiting the Rho pathway strongly suggest that this is one of the mechanisms by which the hydrogel promotes type II phenotype maintenance. These results regarding protein expression strongly suggest that the chemical and biophysical properties of the hydrogel have a considerable impact on the transition from ATII to ATI phenotypes. In conclusion, culturing primary alveolar epithelial cells on lung ECM-derived hydrogels may facilitate the prolonged culturing of these cells, and thus help in the research on lung diseases. 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