A novel human cell culture model to study visceral smooth muscle phenotypic modulation in health and disease

Rianne D. W. Vaes*, Linda van den Berk, Bas Boonen, David P. J. van Dijk, Steven W. M. Olde Damink, Sander S. Rensen

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Adaptation of the smooth muscle cell (SMC) phenotype is essential for homeostasis and is often involved in pathologies of visceral organs (e.g., uterus, bladder, gastrointestinal tract). In vitro studies of the behavior of visceral SMCs under (patho)-physiological conditions are hampered by a spontaneous, uncontrolled phenotypic modulation of visceral SMCs under regular tissue culture conditions. We aimed to develop a new visceral SMC culture model that allows controlled phenotypic modulation. Human uterine SMCs [ULTR and telomerase-immortalized human myometrial cells (hTERT-HM)] were grown to confluency and kept for up to 6 days on regular tissue culture surfaces or basement membrane (BM) matrix-coated surfaces in the presence of 0-10% serum. mRNA and protein expression and localization of SMC-specific phenotype markers and their transcriptional regulators were investigated by quantitative PCR, Western blotting, and immunofluorescence. Maintaining visceral SMCs confluent for 6 days increased alpha-smooth muscle actin (1.9-fold) and smooth muscle protein 22-alpha (3.1-fold), whereas smooth muscle myosin heavy chain was only slightly upregulated (1.3-fold). Culturing on a BM matrix-coated surface further increased these proteins and also markedly promoted mRNA expression of gamma-smooth muscle actin (15.0-fold), smoothelin (3.5-fold), h-caldesmon (5.2-fold), serum response factor (7.6-fold), and myocardin (8.1-fold). Whereas additional serum deprivation only minimally affected contractile markers, platelet-derived growth factor- BB and transforming growth factor beta 1 consistently reduced versus increased their expression. In conclusion, we present a simple and reproducible visceral SMC culture system that allows controlled phenotypic modulation toward both the synthetic and the contractile phenotype. This may greatly facilitate the identification of factors that drive visceral SMC phenotypic changes in health and disease.

Original languageEnglish
Pages (from-to)C598-C607
Number of pages10
JournalAmerican Journal of Physiology-Cell Physiology
Volume315
Issue number4
DOIs
Publication statusPublished - Oct 2018

Keywords

  • basal membrane matrix
  • differentiation
  • phenotypic modulation
  • visceral smooth muscle
  • EXTRACELLULAR-MATRIX PROTEINS
  • GENE-EXPRESSION
  • OUTLET OBSTRUCTION
  • GROWTH
  • MYOCARDIN
  • ALPHA
  • DIFFERENTIATION
  • FIBRONECTIN
  • CONTRACTILE
  • COLLAGEN

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