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Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction.

Justin Rustenhoven1,2, Leon C Smyth1,2, Deidre Jansson1,2

  • 1Centre for Brain Research, Faculty of Medical and Health Sciences, University of Auckland, 85 Park Rd, Grafton, Auckland, 1023, New Zealand.

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|February 24, 2018
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Summary

Culture media significantly impacts human brain pericyte phenotype and function, influencing proliferation, morphology, and inflammatory responses. However, their core neuroinflammatory response remains robust across different culture conditions, vital for blood-brain barrier research.

Keywords:
Blood–brain barrierGrowth factorInflammationMigrationPhagocytosisProliferation

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Immunology

Background:

  • Brain pericytes are crucial for blood-brain barrier (BBB) integrity and central nervous system (CNS) immune responses.
  • In vitro studies of pericytes are common, but their phenotype and function vary based on culture conditions.
  • This study investigates how two distinct culture media affect human brain pericyte phenotype and neuroinflammatory responses.

Purpose of the Study:

  • To determine the impact of two common culture media (DMEM/F12 with high serum and Pericyte Medium with low serum) on human brain pericyte phenotype.
  • To assess how these culture media influence the neuroinflammatory responses of human brain pericytes.
  • To compare the basal and inflammatory characteristics of pericytes cultured in different media.

Main Methods:

  • Human brain pericytes were isolated and cultured in either DMEM/F12 (D-pericytes) or Pericyte Medium (P-pericytes).
  • Phenotypic differences were assessed using immunocytochemistry, qRT-PCR, and EdU incorporation.
  • Neuroinflammatory responses were evaluated via transcription factor induction, chemokine secretion, adhesion molecule expression, migration, and phagocytosis assays.

Main Results:

  • Pericyte Medium (P-pericytes) cultures showed increased proliferation and a distinct bipolar morphology compared to DMEM/F12 (D-pericytes).
  • P-pericytes exhibited altered expression of key pericyte markers (e.g., lower αSMA, higher Col-IV) and enhanced phagocytic and migratory abilities.
  • While nuclear NF-kB translocation occurred in both, P-pericytes showed elevated C/EBPδ and lower ICAM-1 expression.

Conclusions:

  • Culture media significantly alter human brain pericyte phenotype, including morphology, proliferation, and specific marker expression.
  • Despite phenotypic variations, both pericyte culture conditions demonstrated a robust and similar neuroinflammatory response.
  • These findings highlight the plasticity of brain pericytes in vitro but emphasize the resilience of their core inflammatory functions to culture media differences.