A primary human trophoblast model to study the effect of inflammation associated with maternal obesity on regulation of autophagy in the placenta

Bailey Simon, Matthew Bucher, Alina Maloyan

Research output: Contribution to journalArticle

Abstract

Maternal obesity is associated with an increased risk of adverse perinatal outcomes that are likely mediated by compromised placental function that can be attributed to, in part, the dysregulation of autophagy. Aberrant changes in the expression of autophagy regulators in the placentas from obese pregnancies may be regulated by inflammatory processes associated with both obesity and pregnancy. Described here is a protocol for sampling of villous tissue and isolation of villous cytotrophoblasts from the term human placenta for primary cell culture. This is followed by a method for simulating the inflammatory milieu in the obese intrauterine environment by treating primary trophoblasts from lean pregnancies with tumor necrosis factor alpha (TNFα), a proinflammatory cytokine that is elevated in obesity and in pregnancy. Through the implementation of the protocol described here, it is found that exposure to exogenous TNFα regulates the expression of Rubicon, a negative regulator of autophagy, in trophoblasts from lean pregnancies with female fetuses. While a variety of biological factors in the obese intrauterine environment maintain the potential to modulate critical pathways in trophoblasts, this ex vivo system is especially useful for determining if expression patterns observed in vivo in human placentas with maternal obesity are a direct result of TNFα signaling. Ultimately, this approach affords the opportunity to parse out the regulatory and molecular implications of inflammation associated with maternal obesity on autophagy and other critical cellular pathways in trophoblasts that have the potential to impact placental function.

Original languageEnglish (US)
Article numbere56484
JournalJournal of Visualized Experiments
Volume2017
Issue number127
DOIs
StatePublished - Sep 27 2017

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Autophagy
Trophoblasts
Placenta
Tumor Necrosis Factor-alpha
Obesity
Mothers
Inflammation
Pregnancy
Critical Pathways
Biological Factors
Cell culture
Primary Cell Culture
Tissue
Cytokines
Sampling
Fetus

Keywords

  • Autophagy
  • Human placenta
  • Inflammation
  • Issue 127
  • Maternal obesity
  • Medicine
  • Primary cell culture
  • Rubicon
  • Sexual dimorphism
  • Trophoblasts
  • Tumor necrosis factor alpha (TNFα)
  • Villous tissue
  • Western blot

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

A primary human trophoblast model to study the effect of inflammation associated with maternal obesity on regulation of autophagy in the placenta. / Simon, Bailey; Bucher, Matthew; Maloyan, Alina.

In: Journal of Visualized Experiments, Vol. 2017, No. 127, e56484, 27.09.2017.

Research output: Contribution to journalArticle

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