Rat tracheal epithelial cell differentiation in vitro

Kaartinen, Liisa; Nettesheim, Paul; Adler, Kenneth B.; Randell, Scott H.

doi:10.1007/bf02639383

Cited by 127 publications

(115 citation statements)

References 40 publications

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“…Other previously identified factors required for ciliogenesis may also determine ciliated cell commitment. Studies of primary cultured airway epithelial cells have shown that ciliogenesis requires epidermal growth factor, bovine pituitary hormones, cholera toxin, and retinoic acid in the setting of ALI (8,17). The effect of these factors on specific stages of ciliogenesis has not been studied using ultrastructural analysis, and it is unclear if one or more of these factors can initiate ciliogenesis.…”

Section: Discussionmentioning

confidence: 99%

Role of f-box factor foxj1 in differentiation of ciliated airway epithelial cells

You¹,

Huang²,

Richer³

et al. 2004

American Journal of Physiology-Lung Cellular and Molecular Phys

184

176

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Factors required for commitment of an undifferentiated airway epithelial cell to a ciliated cell are unknown. Cell ultrastructure analysis indicates ciliated cell commitment activates a multistage program involving synthesis of cilia precursor proteins and assembly of macromolecular complexes. Foxj1 is an f-box transcription factor expressed in ciliated cells and shown to be required for cilia formation by gene deletion in a mouse model. To identify a specific role for foxj1 in directing the ciliated cell phenotype, we evaluated the capacity of foxj1 to induce ciliogenesis and direct cilia assembly. In a primary culture model of wild-type mouse airway epithelial cells, foxj1 expression preceded the appearance of cilia and in cultured foxj1 null cells cilia did not develop. Delivery of foxj1 to polarized epithelial cell lines and primary cultured alveolar epithelial cells failed to promote ciliogenesis. Similarly, delivery of foxj1 to wild-type airway epithelial cells did not enhance the total number of ciliated cells. In contrast, delivery of foxj1 to null cells resulted in the appearance of cilia. Analysis revealed that, in the absence of foxj1, null cells contained cilia precursor basal bodies, indicating prior commitment to ciliogenesis. However, the basal bodies were disorganized within the apical compartment and failed to dock with the apical membrane. Reconstitution of foxj1 in null cells restored normal basal body organization, resulting in axoneme growth. Thus foxj1 functions in late-stage ciliogenesis to regulate programs promoting basal body docking and axoneme formation in cells previously committed to the ciliated cell phenotype.

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Section: Discussionmentioning

confidence: 99%

Role of f-box factor foxj1 in differentiation of ciliated airway epithelial cells

You¹,

Huang²,

Richer³

et al. 2004

American Journal of Physiology-Lung Cellular and Molecular Phys

184

176

View full text Add to dashboard Cite

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“…The NHBE cells utilized in the present study were cultured at the air/liquid interface, resulting in fully differentiated primary cell cultures that maintained a well documented structure and function similar to the in vivo one (8,(21)(22)(23). This air/liquid methodology to culture airway epithelial cells was developed several years ago to provide an ideal in vitro model system to study mechanisms involved in various cellular processes in airway epithelium.…”

Section: Discussionmentioning

confidence: 99%

MARCKS Protein Is a Key Molecule Regulating Mucin Secretion by Human Airway Epithelial Cells in Vitro

Li¹,

Martin²,

Spizz³

et al. 2001

Journal of Biological Chemistry

Self Cite

158

186

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Hypersecretion of airway mucin characterizes numerous respiratory diseases. Although diverse pathological stimuli can provoke exocytotic release of mucin from secretory cells of the airway epithelium, mechanisms involved remain obscure. This report describes a new paradigm for the intracellular signaling mechanism regulating airway mucin secretion. Direct evidence is provided that the myristoylated alanine-rich C kinase substrate (MARCKS) is a central regulatory molecule linking secretagogue stimulation at the cell surface to mucin granule release by differentiated normal human bronchial epithelial cells in vitro. Down-regulation of MARCKS expression or disruption of MARCKS function in these cells inhibits the secretory response to subsequent stimulation. The intracellular mechanism controlling this secretory process involves cooperative action of two separate protein kinases, protein kinase C and cGMP-dependent protein kinase. Upon stimulation, activated protein kinase C phosphorylates MARCKS, causing translocation of MARCKS from the plasma membrane to the cytoplasm, where it is then dephosphorylated by a protein phosphatase 2A that is activated by cGMP-dependent protein kinase, and associates with both actin and myosin. Dephosphorylated cytoplasmic MARCKS would also be free to interact with mucin granule membranes and thus could link granules to the contractile cytoskeleton, mediating their movement to the cell periphery and subsequent exocytosis. These findings suggest several novel intracellular targets for pharmacological intervention in disorders involving aberrant secretion of respiratory mucin and may relate to other lesions involving exocytosis of membrane-bound granules in various cells and tissues.

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“…Media formulations were modified from previously described methods (14,17,19,(26)(27)(28). Supplements were from Sigma-Aldrich (St. Louis, MO) unless indicated.…”

Section: Methodsmentioning

confidence: 99%

Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population

You

Richer

Huang

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

418

578

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Highly regulated programs for airway epithelial cell proliferation and differentiation during development and repair are often disrupted in disease. These processes have been studied in mouse models; however, it is difficult to isolate and identify epithelial cell-specific responses in vivo. To investigate these processes in vitro, we characterized a model for primary culture of mouse tracheal epithelial cells. Small numbers of cells seeded at low density (7.5 × 104 cells/cm2) rapidly proliferated and became polarized. Subsequently, supplemented media and air-liquid interface conditions resulted in development of highly differentiated epithelia composed of ciliated and nonciliated cells with gene expression characteristic of native airways. Genetically altered or injured mouse tracheal epithelial cells also reflected in vivo patterns of airway epithelial cell gene expression. Passage of cells resulted in continued proliferation but limited differentiation after the first passage, suggesting that transit-amplifying cell populations were present but with independent programs for proliferation and differentiation. This approach provides a high-fidelity in vitro model for evaluation of gene regulation and expression in mouse airway epithelial cells.

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Rat tracheal epithelial cell differentiation in vitro

Cited by 127 publications

References 40 publications

Role of f-box factor foxj1 in differentiation of ciliated airway epithelial cells

Role of f-box factor foxj1 in differentiation of ciliated airway epithelial cells

MARCKS Protein Is a Key Molecule Regulating Mucin Secretion by Human Airway Epithelial Cells in Vitro

Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population

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