Primary culture of rat alveolar type II cells on floating collagen membranes

Geppert, E; Williams, Mary C.; Mason, Robert J.

doi:10.1016/0014-4827(80)90072-5

Cited by 65 publications

(21 citation statements)

References 16 publications

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“…We rocked the cultures so that the epithelial cells would be exposed briefly to the gas phase as occurs in the lung in vivo. The substratum on which the type II cells were cultured was a mixture of 20% (by volume) EHS matrix (Matrigel) and 80% rat-tail collagen that we prepared (16). On gels containing a very high percentage of EHS, type II cells formed spherules that trapped the apical secretions, and secretion of SP-A and SP-D into the medium decreased substantially.…”

Section: Resultsmentioning

confidence: 99%

“…Maintaining type II cells on tissue culture plastic leads to cell spreading and dedifferentiation as determined by decreased expression of surfactant protein mRNAs and altered patterns of phospholipid biosynthesis. Improvement in functional differentiation has been achieved by culturing type II cells on contracted collagen gels, Engelbreth-Holm-Swarm (EHS) tumor basement-membrane matrix, human amnion, and filters coated with extracellular matrix (9,16,(42)(43)(44)52). Substrata rich in laminin maintain the cells in a more cuboidal shape and are beneficial, whereas substrata rich in fibronectin cause the cells to flatten and dedifferentiate (5,36).…”

mentioning

confidence: 99%

“…A cuboidal shape and thereby a permissive cytoarchitecture appears to be a prerequisite for maintaining type II cell differentiation and surfactant protein mRNA stability (45). This cell shape can be achieved by culturing type II cells on EHS gels where the cells form spherules or by having a substratum such as a floating collagen gel that allows the cells to contract the gel and assume a native cuboidal shape (16,(42)(43)(44). Neither of these two strategies requires cell growth to increase the number of cells per surface area to maintain a cuboidal shape.…”

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confidence: 99%

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Maintenance of surfactant protein A and D secretion by rat alveolar type II cells in vitro

Mason

Lewis

Edeen

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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Secretion of surfactant proteins A and D (SP-A and SP-D) has been difficult to study in vitro because a culture system for maintaining surfactant secretion has been difficult to establish. We evaluated several growth factors, corticosteroids, rat serum, and a fibroblast feeder layer for the ability to produce and maintain a polarized epithelium of type II cells that secretes SP-A and SP-D into the apical medium. Type II cells were plated on a filter insert coated with an extracellular matrix and were cultured at an air-liquid interface. Keratinocyte growth factor (KGF) stimulated type II cell proliferation and secretion of SP-A and SP-D more than fibroblast growth factor-10 (FGF-10), hepatocyte growth factor (HGF), or heparin-binding epidermal-like growth factor (HB-EGF). Cells cultured in the presence of KGF and rat serum with or without fibroblasts had high surfactant protein mRNA levels and exhibited a high level of SP-A and SP-D secretion. Dexamethasone inhibited type II cell proliferation but increased expression of SP-B. In the presence of KGF, rat serum, and dexamethasone, the mRNAs for the surfactant proteins were maintained at high levels. Secretion of SP-A and SP-D was found to be independent of phospholipid secretion.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Maintenance of surfactant protein A and D secretion by rat alveolar type II cells in vitro

Mason

Lewis

Edeen

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Type II pneumocytes have been shown to retain their cuboidal morphology when cultured on various ECM substrates, in contrast to the loss of morphology and spreading seen on tissue culture plastic. In previous studies the substrates used were pure single components of ECM (9,16) or complete ECM from cells of known origin (15)(16)(17)(18)(19). However, in this study the exact source and composition of the extracellular matrix is unclear.…”

Section: Discussionmentioning

confidence: 89%

“…Neither do they divide in isolated cell culture (13,14). The normal morphology of Type II cells in vitro can be prolonged by plating onto exogenous extracellular matrices (ECM), such as laminin (15), collagen gels (16) or basement membrane derived from cultured cells (15,17), and other sources (18,19). Furthermore, macrophage-or fibroblast-derived factors are thought to influence the differentiated function of Type H cells in vitro (17,20).…”

Section: Introductionmentioning

confidence: 99%

Type II Pneumocytes in Mixed Cell Culture of Human Lung: A Light and Electron Microscopic Study

Bingle¹,

Bull²,

Fox³

et al. 1990

Environmental Health Perspectives

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An acellular human amnionic membrane model for in vitro culture of type ii pneumocytes: The role of the basement membrane in cell morphology and function

Lwebuga‐Mukasa

Thulin

Madri

et al. 1984

Journal Cellular Physiology

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To determine whether a preformed basement membrane contributes to the maintenance of morphology and function of type II pneumocytes, we cultured isolated adult rat type II pneumocytes on the basement membrane and stromal surfaces of an acellular human amnionic membrane and on plastic. The presence of lamellar bodies on transmission electron microscopy and epithelial morphology in culture and a characteristic phospholipid profile after incubation with 3H-acetate identified the cells as type II. When type II cells were cultured on a preexisting basement membrane, they formed a well-organized monolayer with polarity, centrally located surface microvilli, and more basally located nuclei. Individual cells maintained a cuboidal morphology for 8-10 days. Intracellularly, there were numerous mitochondria, endoplasmic reticulum (ER), and lamellar bodies. The cells secreted a new basal lamina of their own. When cultured on the stromal side of the amnion, the cells became flattened within 48-60 hours, formed small lamellar bodies, and had scanty surface microvilli; they formed clumps and appeared less ordered. These cells did not secrete a visible basement membrane, and the majority detached from the stromal surface after 7-8 days in culture. In addition, culture on the basement membrane aspect of the amnion prevented the rapid decline in the percentage of 3H-acetate label incorporated in phosphatidylcholine after 72 hours of culture. We conclude that a preformed basement membrane influences the function and morphology of type II pneumocytes, organizes them into a monolayer in culture, and influences deposition of a visible basal lamina. Thus, the acellular human amnion provides an excellent model for the systematic study of basement membrane influence on the biology and pathology of these cells.

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Primary culture of rat alveolar type II cells on floating collagen membranes

Cited by 65 publications

References 16 publications

Maintenance of surfactant protein A and D secretion by rat alveolar type II cells in vitro

Maintenance of surfactant protein A and D secretion by rat alveolar type II cells in vitro

Type II Pneumocytes in Mixed Cell Culture of Human Lung: A Light and Electron Microscopic Study

An acellular human amnionic membrane model for in vitro culture of type ii pneumocytes: The role of the basement membrane in cell morphology and function

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