Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs
Summary: Currently, the only approach for obtaining alveolar epithelial type I (AT1) cells is through the differentiation of cultured AT2 cells, which has several limitations. Here, we present a protocol to prepare AT1 cells directly from mouse lungs. We detail the steps for dissociating epithelial...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-12-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672400649X |
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| author | Lulu Huang Amy Tang Bojing Shao |
| author_facet | Lulu Huang Amy Tang Bojing Shao |
| author_sort | Lulu Huang |
| collection | DOAJ |
| description | Summary: Currently, the only approach for obtaining alveolar epithelial type I (AT1) cells is through the differentiation of cultured AT2 cells, which has several limitations. Here, we present a protocol to prepare AT1 cells directly from mouse lungs. We detail the steps for dissociating epithelial cells from the alveoli, followed by the purification of AT1 cells. Additionally, we describe how to assess the purity of AT1 cells using immunofluorescence staining and flow cytometry, along with approaches for verifying their functions. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-3b5fbae1233c4a79a6283fcf85f7867e |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-3b5fbae1233c4a79a6283fcf85f7867e2025-08-20T01:59:35ZengElsevierSTAR Protocols2666-16672024-12-015410348410.1016/j.xpro.2024.103484Protocol for preparation of primary alveolar epithelial type I cells from mouse lungsLulu Huang0Amy Tang1Bojing Shao2Laboratory of Vascular Inflammation and Thrombosis Research, Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10065, USALaboratory of Vascular Inflammation and Thrombosis Research, Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10065, USALaboratory of Vascular Inflammation and Thrombosis Research, Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10065, USA; Corresponding authorSummary: Currently, the only approach for obtaining alveolar epithelial type I (AT1) cells is through the differentiation of cultured AT2 cells, which has several limitations. Here, we present a protocol to prepare AT1 cells directly from mouse lungs. We detail the steps for dissociating epithelial cells from the alveoli, followed by the purification of AT1 cells. Additionally, we describe how to assess the purity of AT1 cells using immunofluorescence staining and flow cytometry, along with approaches for verifying their functions. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672400649Xcell biologycell culturecell isolationcell separation/fractionationmass cytometry |
| spellingShingle | Lulu Huang Amy Tang Bojing Shao Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs STAR Protocols cell biology cell culture cell isolation cell separation/fractionation mass cytometry |
| title | Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs |
| title_full | Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs |
| title_fullStr | Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs |
| title_full_unstemmed | Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs |
| title_short | Protocol for preparation of primary alveolar epithelial type I cells from mouse lungs |
| title_sort | protocol for preparation of primary alveolar epithelial type i cells from mouse lungs |
| topic | cell biology cell culture cell isolation cell separation/fractionation mass cytometry |
| url | http://www.sciencedirect.com/science/article/pii/S266616672400649X |
| work_keys_str_mv | AT luluhuang protocolforpreparationofprimaryalveolarepithelialtypeicellsfrommouselungs AT amytang protocolforpreparationofprimaryalveolarepithelialtypeicellsfrommouselungs AT bojingshao protocolforpreparationofprimaryalveolarepithelialtypeicellsfrommouselungs |