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Isolation and culture of mouse alveolar type II cells to study type II to type I cell differentiation.

ABSTRACT: Alveolar type II cells (AT2) are stem cells of lung epithelium. To study these cells, we developed protocols to isolate primary AT2 and test the effects of potential niche factors in regulating the AT2 progenitor functions. AT2 freshly isolated from mouse lungs are grown in 2D or 3D culture. AT2 are able to differentiate into alveolar type I cells (AT1) in these conditions. S1P or an inhibitor of S1P signaling is added in culture to modulate AT2 to AT1 transition. For complete details on the use and execution of this protocol, please refer to Chen et al. (2020).

SUBMITTER: Chen Q

PROVIDER: S-EPMC7788236 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Isolation and culture of mouse alveolar type II cells to study type II to type I cell differentiation.

Chen Qian Q Liu Yuru Y

STAR protocols 20201231 1

Alveolar type II cells (AT2) are stem cells of lung epithelium. To study these cells, we developed protocols to isolate primary AT2 and test the effects of potential niche factors in regulating the AT2 progenitor functions. AT2 freshly isolated from mouse lungs are grown in 2D or 3D culture. AT2 are able to differentiate into alveolar type I cells (AT1) in these conditions. S1P or an inhibitor of S1P signaling is added in culture to modulate AT2 to AT1 transition. For complete details on the use ...[more]

PMID: 33437966

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ErbB4 alternative splicing mediates fetal mouse alveolar type II cell differentiation in vitro.

Project description:BackgroundAlternative splicing (AS) creates different protein isoforms, an important mechanism regulating cell-specific function. Little is known about AS in lung development, particularly in alveolar type II (ATII) cells. ErbB4 receptor isoforms Jma and Jmb have significant and opposing functions in the brain, heart, and lung development and/or disease. However, the regulators of ErbB4 AS are unknown. ErbB4 AS regulators in fetal mouse ATII cells control its function in ATII cell maturation.MethodsCandidate ErbB4 AS regulators were found using in silico analysis. Their developmental expression was studied in fetal mouse ATII cells. The effects of splice factor downregulation and upregulation on ATII cell maturation were analyzed.ResultsErbB4-Jma increased significantly in ATII cells after gestation E16.5. In silico analysis found four candidate splice factors: FOX2, CUG/CELF1, TIAR, and HUB. Fetal ATII cells expressed these factors in distinct developmental profiles. HUB downregulation in E17.5 ATII cells increased Jma isoform levels and Sftpb gene expression and decreased Jmb. HUB overexpression decreased Jma and Sftpb.ConclusionsErbB4 AS is developmentally controlled by HUB in fetal ATII cells, promoting ATII differentiation. Regulated AS expression during ATII cell differentiation suggests novel therapeutic strategies to approach human disease.ImpactAlternative splicing (AS) of the ErbB4 receptor, involving mutually exclusive exon inclusion, creates Jma and Jmb isoforms with distinct differences in receptor processing and function. The Jma isoform of ErbB4 promotes differentiation of fetal lung alveolar type II cells. The AS is mediated in part by the RNA-binding protein HUB. The molecular mechanism of AS for ErbB4 has not been previously described. The regulation of ErbB4 AS has important implications in the development of organs, such as the lung, brain, and heart, and for disease, including cancer.

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Human alveolar epithelial type II cells in primary culture.

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Silver nanowire interactions with primary human alveolar type-II epithelial cell secretions: contrasting bioreactivity with human alveolar type-I and type-II epithelial cells.

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Dissecting the Niche for Alveolar Type II Cells With Alveolar Organoids.

Project description: Not available

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