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Rotational co‐culture of clonal β ‐cells with endothelial cells: effect of PPAR‐ γ agonism in vitro on insulin and VEGF secretion
Author(s) -
Paget M. B.,
Murray H. E.,
Bailey C. J.,
Flatt P. R.,
Downing R.
Publication year - 2011
Publication title -
diabetes, obesity and metabolism
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.445
H-Index - 128
eISSN - 1463-1326
pISSN - 1462-8902
DOI - 10.1111/j.1463-1326.2011.01392.x
Subject(s) - medicine , endocrinology , insulin , umbilical vein , secretion , rosiglitazone , vascular endothelial growth factor , biology , receptor , in vitro , chemistry , biochemistry , vegf receptors
Aim: Delayed graft revascularization impedes the success of human islet transplantation. This study utilized rotational co‐culture of insulin secreting β ‐cells with human umbilical vein endothelial cells (HUVECs) and a peroxisome proliferator‐activated receptor gamma (PPAR‐ γ ) agonist to promote insulin and vascular endothelial growth factor (VEGF) secretory function. Methods: Clonal BRIN‐BD11 (D11) cells were maintained in static culture (SC) and rotational culture (RC) ± HUVEC and ± the TZD (thiazolidinedione) rosiglitazone (10 mmol/l) as a specific PPAR‐ γ agonist. HUVECs were cultured in SC and RC ± D11 and ± TZD. D11 insulin secretion was induced by static incubation with low glucose (1.67 mmol/l), high glucose (16.7 mmol/l) and high glucose with 10 mmol/l theophylline (G+T) and assessed by enzyme‐linked immunosorbent assay (ELISA). HUVEC proliferation was determined by ATP luminescence, whereas VEGF secretion was quantified by ELISA. Co‐cultured cells were characterized by immunostaining for insulin and CD31. Results: D11 SC and RC showed enhanced insulin secretion in response to 16.7 mmol/l and G+T (p < 0.01); without significant alteration by the TZD. Co‐culture with HUVEC in SC and RC also increased D11 insulin secretion when challenged with 16.7 mmol/l and G+T (p < 0.01), and this was slightly enhanced by the TZD. The presence of HUVEC increased D11 SC and RC insulin secretion in response to high glucose and G+T, respectively (p < 0.01). Addition of the TZD increased SC and RC HUVEC ATP content (p < 0.01) and VEGF production (p < 0.01) in the presence and absence of D11 cells. Conclusions: Rotational co‐culture of insulin secreting cells with endothelial cells, and exposure to a PPAR‐ γ agonist may improve the prospects for graft revascularization and function after implantation.