Preservation of glucose responsiveness in human islets maintained in a rotational cell culture system.
The diminution of glucose responsiveness in isolated human islets maintained under conventional static culture (CSC) conditions represents a major limitation for the long-term storage of islet tissue and precludes extensive study of beta (beta)-cell biology. In the present investigation, we examined the effect of culturing primary human islets in a rotational cell culture system (RCCS) to determine its' ability to sustain both the structural integrity and functional viability of these fragile cell constructs. Over a 10-day culture period both structural integrity and glucose-stimulated insulin release (GSIR) were preserved in islets maintained within the RCCS whilst those held under CSC conditions exhibited progressive fragmentation and rapid loss of secretory function. In addition, intentionally dissociated islet cells maintained within the RCCS demonstrated the ability to re-aggregate and form tight islet-like structures with enhanced secretory capacity compared to whole islets maintained in static culture. These findings suggest a novel use for the RCCS and illustrate its potential as an experimental tool for in vitro study of human islet/beta-cell physiology.
Murray
Hilary E
HE
Islet Research Laboratory, Worcestershire Clinical Research Unit, Worcestershire Acute Hospitals NHS Trust, Newtown Road, Worcester WR5 1HN, UK. hilary.murray@worcsacute.wmids.nhs.uk
Paget
Michelle B
MB
Downing
Richard
R
D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010477:Perfusion; D012399:Rotation; D013997:Time Factors
D007328:Insulin; D005947:Glucose