title: Sustained insulin secretory response in human islets co-cultured with pancreatic duct-derived epithelial cells within a rotational cell culture system.
abstract: OBJECTIVE
  Loss of the trophic support provided by surrounding non-endocrine pancreatic cell populations underlies the decline in beta cell mass and insulin secretory function observed in human islets following isolation and culture. This study sought to determine whether restoration of regulatory influences mediated by ductal epithelial cells promotes sustained beta cell function in vitro.
  
  
  METHODS
  Human islets were isolated according to existing protocols. Ductal epithelial cells were harvested from the exocrine tissue remaining after islet isolation, expanded in monolayer culture and characterised using fluorescence immunocytochemistry. The two cell types were co-cultured under conventional static culture conditions or within a rotational cell culture system. The effect of co-culture on islet structural integrity, beta cell mass and insulin secretory capacity was observed for 10 days following isolation.
  
  
  RESULTS
  Human islets maintained under conventional culture conditions exhibited a characteristic loss in structural integrity and functional viability as indicated by a diminution of glucose responsiveness. By contrast, co-culture of islets with ductal epithelial cells led to preserved islet morphology and sustained beta cell function, most evident in co-cultures held within the rotational cell culture system, which showed a significantly (p < 0.05) greater insulin secretory response to elevated glucose compared with control islets. Similarly, insulin/protein ratio data suggested that the presence of ductal epithelial cells is beneficial for the maintenance of beta cell mass.
  
  
  CONCLUSIONS
  The data indicate a supportive role for ductal epithelial cells in islet viability. Further characterisation of the regulatory influences may lead to novel strategies to improve long-term beta cell function both in vitro and following islet transplantation.
authors.affiliation: The Islet Research Laboratory, Worcestershire Clinical Research Unit, Worcestershire Acute Hospitals NHS Trust, Newtown Road, Worcester, WR5 1HN, UK. hilary.murray@worcsacute.nhs.uk
authors.forename: H E
authors.initials: HE
authors.lastname: Murray
authors.forename: M B
authors.initials: MB
authors.lastname: Paget
authors.forename: C J
authors.initials: CJ
authors.lastname: Bailey
authors.forename: R
authors.initials: R
authors.lastname: Downing
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D018920:Coculture Techniques / Q000379:methods; D004847:Epithelial Cells / Q000166:cytology*; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism*; D010183:Pancreatic Ducts / Q000166:cytology*
chemical_list: D007328:Insulin; D005947:Glucose