Influence of Culture Substrates on Morphology and Function of Pulmonary Alveolar Cells In Vitro

Biomolecules. 2021 Apr 30;11(5):675. doi: 10.3390/biom11050675.

Abstract

Cell's microenvironment has been shown to exert influence on cell behavior. In particular, matrix-cell interactions strongly impact cell morphology and function. The purpose of this study was to analyze the influence of different culture substrate materials on phenotype and functional properties of lung epithelial adenocarcinoma (A549) cells. A549 cells were seeded onto two different biocompatible, commercially available substrates: a polyester coverslip (Thermanox™ Coverslips), that was used as cell culture plate control, and a polydimethylsiloxane membrane (PDMS, Elastosil® Film) investigated in this study as alternative material for A549 cells culture. The two substrates influenced cell morphology and the actin cytoskeleton organization. Further, the Yes-associated protein (YAP) and its transcriptional coactivator PDZ-binding motif (TAZ) were translocated to the nucleus in A549 cells cultured on polyester substrate, yet it remained mostly cytosolic in cells on PDMS substrate. By SEM analysis, we observed that cells grown on Elastosil® Film maintained an alveolar Type II cell morphology. Immunofluorescence staining for surfactant-C revealing a high expression of surfactant-C in cells cultured on Elastosil® Film, but not in cells cultured on Thermanox™ Coverslips. A549 cells grown onto Elastosil® Film exhibited morphology and functionality that suggest retainment of alveolar epithelial Type II phenotype, while A549 cells grown onto conventional plastic substrates acquired an alveolar Type I phenotype.

Keywords: cell function; cell morphology; lung epithelial cells; mechanobiology; substrate properties.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Adaptor Proteins, Signal Transducing / biosynthesis
  • Alveolar Epithelial Cells / cytology*
  • Alveolar Epithelial Cells / drug effects*
  • Cell Culture Techniques / methods
  • Cell Line, Tumor
  • Culture Media
  • Dimethylpolysiloxanes / pharmacology*
  • Humans
  • Lipopeptides / biosynthesis
  • Lung / cytology
  • Lung / drug effects
  • Microscopy, Electrochemical, Scanning
  • Peptides, Cyclic / biosynthesis
  • Polyesters / pharmacology*
  • Pulmonary Alveoli / cytology*
  • Pulmonary Alveoli / drug effects*
  • Transcription Factors / biosynthesis
  • YAP-Signaling Proteins

Substances

  • Adaptor Proteins, Signal Transducing
  • Culture Media
  • Dimethylpolysiloxanes
  • Lipopeptides
  • Peptides, Cyclic
  • Polyesters
  • Transcription Factors
  • YAP-Signaling Proteins
  • YAP1 protein, human
  • surfactin peptide
  • baysilon