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TP1-4 In vitro induced cytokine response of astrocytes modelling conditions in human traumatic brain injury
  1. EP Thelin1,
  2. CE Hall2,
  3. A Frostell3,
  4. T Tajsic1,
  5. KLH Carpenter1,
  6. PJA Hutchinson1,
  7. R Patani2,
  8. A Helmy1
  1. 1University of Cambridge, Cambridge, UK
  2. 2University College London, London, UK
  3. 3Karolinska Institutet, Stockholm, Sweden


Objectives The objective of this study was to investigate how in vitro astrocyte cultures respond to cytokine pro- and anti-inflammatory cytokine concentrations, corresponding to those seen in the aftermath of human TBI, by analysing downstream cytokine generation.

Design In vitro study.

Subjects Human induced pluripotent stem cells (iPSC)-derived astrocytes.

Methods The astrocytes were exposed to levels of TNF (1–10,000 pg/ml), IL-6 (100–1,000,000 pg/ml), Interleukin-1β (IL-1β, 1–10,000 pg/ml), Interleukin-4 (IL-4, 1–10,000 pg/ml) and Interleukin-10 (IL-10, 1–10,000 pg/ml). Following 24, 48 and 72 hours, culture supernatant was extracted and analysed using a human cytokine/chemokine 39-plex luminex assay (ThermoFisher).

Results The astrocyte secretome revealed concentration-, time- or concentration*time-dependent production of downstream cytokines (12, 8 and 2 cytokines, respectively p<0.05). IL-1β and TNF exposure generated the most downstream cytokine production, while IL-6, IL-4 and IL-10 did not generally induce a robust response.

Conclusions iPSC-derived astrocytes exposed to cytokine concentrations reflecting those in TBI generate an increased downstream cytokine production, especially when exposed to IL-1β and TNF. This is in contrast to our previous work on neuronal cultures where IL-1β only produced a few down-stream cytokines.1 More work is needed to better understand how different cells in the CNS respond to the neuroinflammatory milieu after TBI alone and in combination.

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