Co-culture of insulin producing human EndoC-βH1 cells with boundary cap neural crest stem cells protects partially against cytokine-induced cell death
(English)Manuscript (preprint) (Other academic)
We have recently observed that co-culture of mouse and rat beta-cells with mouse boundary cap neural crest stem cells (bNCSCs) protected against inflammatory cytokine-induced cell death, possibly via direct cadherin-mediated cell-to-cell junctions. However, it has not been addressed whether also human beta-cells can be protected via this strategy. The aim of this investigation was therefore to study the effect of bNCSC co-culture with insulin producing human EndoC-βH1 cells on cytokine-induced cell death. For this purpose GFP-positive bNCSCs were cultured together with GFP-negative EndoC-βH1 cells in the presence of the cytokines IL-1b (50 U/ml) and IFN-g (1000 U/ml). Cells were then stained with propidium iodide and trypsinized for flow cytometry analysis. Analysis of propidium iodide fluorescence in GFP-positive and GFP-negative cells revealed that EndoC-βH1 cells died to a lower extent when co-cultured with bNCSCs than when cultured without bNCSCs. We also observed that EndoC-βH1 cells formed N-cadherin, but not E-cadherin junctions with the bNCSCs. The bNCSC cell population contained a large proportion of beta-tubulin expressing cells indicating neuronal differentiation. A protective function of the N-cadherin junctions was suggested by one experiment in which a neutralizing N-cadherin antibody counteracted the effect of co-culture. We conclude that the interaction between human insulin producing cells and bNCSCs results in a lowered susceptibility of insulin producing cells to pro-inflammatory cytokines in vitro.
Medical and Health Sciences
Research subject Biomedical Laboratory Science
IdentifiersURN: urn:nbn:se:uu:diva-233153OAI: oai:DiVA.org:uu-233153DiVA: diva2:750720