An Air–Liquid Interface Promotes the Differentiation of Gastric Surface Mucous Cells (GSM06) in Culture

Ootani, Akifumi; Toda, Shuji; Fujimoto, Kazuma; Sugihara, Hajime

doi:10.1006/bbrc.2000.2673

Cited by 42 publications

(52 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…I AL cultures are abundantly used for various cell types that may have constitutive or acute contact with air; e.g., bronchial and tracheal airway epithelial cells (33), nasal mucosal cells (11), middle ear epithelial cells (42), gastric surface mucus cells (38), various cell lines representing any part of the respiratory tract (21), lung slices (46), but also skin fibroblasts and epidermal keratinocytes (47), to give some examples. The techniques used range from simple hydrogels to filter-based systems up to highly specialized devices (27) and are applied for long-term studies in most cases.…”

Section: )mentioning

confidence: 99%

Interfacial sensing by alveolar type II cells: a new concept in lung physiology?

Ravasio

Hobi²,

Bertocchi³

et al. 2011

American Journal of Physiology-Cell Physiology

View full text Add to dashboard Cite

Alveolar type II (AT II) cells are in close contact with an air-liquid interface (IAL). This contact may be of considerable physiological relevance; however, no data exist to provide a satisfying description of this specific microenvironment. This is mainly due to the experimental difficulty to manipulate and analyze cell-air contacts in a specific way. Therefore, we designed assays to quantify cell viability, Ca 2ϩ changes, and exocytosis in the course of interface contact and miniaturized IAL devices for direct, subcellular, and real-time analyses of cell-interface interactions by fluorescence microscopy or interferometry. The studies demonstrated that the sole presence of an IAL is not sensed by the cells. However, when AT II cells are forced into closer contact with it, they respond promptly with sustained Ca 2ϩ signals and surfactant exocytosis before the occurrence of irreversible cell damage. This points to a paradoxical situation: a potential threat and potent stimulus for the cells. Furthermore, we found that the signalling mechanism underlying sensation of an I AL can be sufficiently explained by mechanical forces. These results demonstrate that the IAL itself can play a major, although so-far neglected, role in lung physiology, particularly in the regulatory mechanisms related with surfactant homeostasis. Moreover, they also support a general new concept of mechanosensation in the lung. mechanical stress; pneumocytes; strain; stretch; surfactant THE ALVEOLAR EPITHELIUM is covered by a thin and continuous layer of water (5). This aqueous layer, referred to as hypophase or alveolar lining fluid (ALF), introduces a considerable physical instability to the millions of alveolar invaginations, tending on overall to force the air out of the lungs. As a consequence, the great alveolar corner cells (or alveolar type II cells; AT II) synthesize and release surfactant into the ALF, from where it transits to the air-liquid interface (I AL ) and creates a highly surface-active coat (40). The importance of this protective coat is best demonstrated in the premature infant lung, where deficiency causes alveolar collapse with life-threatening consequences unless medicated by surfactant administration strategies (17).An important and almost unique aspect of the AT II cellspecific microenvironment is the presence of air. Not astonishingly, therefore, studies introducing such an I AL in AT II cell culture systems reported dramatic effects on cell function, morphology, protein expression, and ion channel activities (13,29). In general, the AT II cells have a higher phenotypic stability and seem to preserve characteristic cell functions for longer periods than compared with standard culture conditions. In particular, biosynthesis and secretion of surfactant is stable over several weeks. Furthermore, reculturing with exposure to air reverses the loss of differentiated AT II cell phenotype observed in submerged cultures (13). This effect of an I AL is indeed remarkable, though not readily intelligible. In particular, the phy...

show abstract

Section: )mentioning

confidence: 99%

Interfacial sensing by alveolar type II cells: a new concept in lung physiology?

Ravasio

Hobi²,

Bertocchi³

et al. 2011

American Journal of Physiology-Cell Physiology

View full text Add to dashboard Cite

show abstract

“…So far, the design of such a robust in vitro system has not been reported for the study of H. pylori-epithelial cell interactions. Two major difficulties have to be overcome: 1) gastric cell lines cannot be grown as polarized epithelia mimicking the gastric mucosa (5-8), 2) or if so (9) do not stand the microaerophilic conditions needed for the particular metabolism of H. pylori. The low oxygen conditions prevailing in the gastric environment are important in the activation of H. pylori virulence genes (10).…”

mentioning

confidence: 99%

Microaerophilic Conditions Permit to Mimic in VitroEvents Occurring during in Vivo Helicobacter pylori Infection and to Identify Rho/Ras-associated Proteins in Cellular Signaling

Cottet

Corthésy–Theulaz

Spertini

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

Molecular dissection of the mechanisms underlyingHelicobacter pylori infection suffers from the lack of in vitro systems mimicking in vivo observations. A system was developed whereby human epithelial cells (Caco-2) grown as polarized monolayers and bacteria can communicate with each other under culture conditions optimal for each partner. Caco-2 cells grown on filter supports were inserted in a vertical position into diffusion chambers equilibrated with air and 5% CO 2 at their basolateral surface (aerophilic conditions) and 5% CO 2 , 5% O 2 , 90% N 2 (microaerophilic conditions) in the apical compartment. Remarkably, the epithelial polarized layer was stable under these asymmetric culture conditions for at least 24 h, and the presence of Caco-2 cells was necessary to maintain H. pylori growth. In contrast to previous studies conducted with non-polarized Caco-2 cells and other cell lines kept under aerophilic conditions, we found H. pylori-dependent stimulation of cytokine secretion (MCP-1 (monocyte chemoattractant protein-1), GRO-␣ (growth-regulated oncogene-␣), RANTES (regulated on activation normal T cell expressed and secreted)). This correlated with nuclear translocation of NF-B p50 and p65 subunits. Tyrosine phosphorylation of nine cellular proteins was induced or enhanced; we identified p120RasGAP , p190 RhoGAP , p62dok (downstream of tyrosine kinases), and cortactin as H. pylori-inducible targets. Moreover, reduction of H. pylori urease expression was observed in adherent bacteria as compared with bacteria in suspension. In addition to mimicking several observations seen in the inflamed gastric mucosa, the novel in vitro system was allowed to underscore complex cellular events not seen in classical in vitro analyses of microaerophilic bacteria-epithelial cell cross-talk.

show abstract

“…We established gastric surface mucous cell lines (GSM06 and GSM10) (Sugiyama et al, 1993) and tracheal epithelial cell lines (TM01 and TM02-3) (Sugiyama et al, 1998) from primary cultures of gastric and tracheal epithelial cells, respectively, of the transgenic mice harboring the tsSV40 large T-antigen gene. Interestingly, these cell lines have specific functions; GSM06 and GSM10 cells produce mucous layer on the cell surface (Dohi et al, 1996;Goso et al, 1997;Ootani et al, 2000;Sugiyama et al, 1993, and TM01 and TM02-3 cells produce high-molecularweight glycoconjugates (Sugiyama et al, 1998).…”

mentioning

confidence: 99%

Establishment and Characterization of a Colonic Epithelial Cell Line MCE301 from Transgenic Mice Harboring Temperature-Sensitive Simian Virus 40 Large T-Antigen Gene.

Tabuchi

Ohta

Arai³

et al. 2000

Cell Struct. Funct.

View full text Add to dashboard Cite

ABSTRACT. We produced an immortalized colonic epithelial cell line, MCE301, using fetal mice transgenic for the temperature-sensitive simian virus 40 large T-antigen gene. MCE301 cells showed epithelial-like morphology and maintained tight connections with neighboring cells. The cells grew at a permissive temperature (33°°°°C), but the growth of the cells was significantly prevented at the nonpermissive temperature (39°°°°C). The cells expressed large T-antigen at 33°°°°C but not at 39°°°°C. MCE301 cells were not transformed, as judged by the absence of anchorage-independent growth in soft agar gel and lack of tumor formation in nude mice. Electron microscopic studies showed that the cells formed microvilli-like structures on the cell surface and junctional complexes such as tight junctions and desmosomes between the cells. The cells expressed cytosketal (acidic cytokeratins and actin), basement membrane (laminin and collagen type IV) and junctional complex proteins (ZO-1 and desmoplakin I + + + + II), as judged by specific antibodies. Fetal bovine serum, epidermal growth factor, insulin-like growth factor and insulin significantly increased the cell growth at 33°°°°C. Moreover, MCE301 cells expressed colonic mucin Muc2 mRNA as demonstrated by reverse transcriptase-polymerase chain reaction, indicating that the cells originate from mucus-secreting cells. Alkaline phosphatase, a brush border-associated enzyme, was detected in the cells. Sodium butyrate (2 mM), an inducer of cellular differentiation, markedly elevated alkaline phosphatase activity. Thus, the present mouse colonic epithelial cell line MCE301 possessing these unique characteristics should provide a useful in vitro model of colonic epithelium.

show abstract

An Air–Liquid Interface Promotes the Differentiation of Gastric Surface Mucous Cells (GSM06) in Culture

Cited by 42 publications

References 29 publications

Interfacial sensing by alveolar type II cells: a new concept in lung physiology?

Interfacial sensing by alveolar type II cells: a new concept in lung physiology?

Microaerophilic Conditions Permit to Mimic in VitroEvents Occurring during in Vivo Helicobacter pylori Infection and to Identify Rho/Ras-associated Proteins in Cellular Signaling

Establishment and Characterization of a Colonic Epithelial Cell Line MCE301 from Transgenic Mice Harboring Temperature-Sensitive Simian Virus 40 Large T-Antigen Gene.

Contact Info

Product

Resources

About