Evaluation of human islet-specific functional quality cultured on different gas-permeable membranes

Transplant Proc. 2008 Mar;40(2):401-2. doi: 10.1016/j.transproceed.2008.01.036.


The aim of this study was to investigate different gas-permeable membranes for culturing human islets. Dynamic insulin release was used to assess islet functional quality. Islets isolated from cadaveric pancreata (n = 8) using standard isolation methods were stained with dithizone, counted, and cultured on five different commercially available medical-grade membranes reported to have high permeability to O2, CO2, and other gases. Fraction 1 (20,000 islet equivalents [IEQ] purity >70%; viability >85%) was cultured using serum-free medium in nonadherence tissue culture flasks (group I) and custom-made chambers with membranes (group II). Each vessel contained 5000 IEQ at a density of 30 IEQ/cm2 and 69 IEQ/cm2 for groups I and II, respectively. Islets were cultured for 48 to 90 hours at 37 degrees C in 5% CO2. In vitro dynamic insulin response to low glucose (3 mmol/L), high glucose (16.7 mmol/L), and 25 mmol/L KCI was measured. Stimulation indices were calculated by dividing average of initial response over basal insulin release; basal insulin release defined as average of the first seven values. Islets cultured on MG7 (n = 3) showed a higher stimulation index (3.49 +/- 0.37) compared with flasks (2.44 +/- 0.22), indicating better specific functional quality. Islets cultured on other membranes proved to show similar or worse functional quality than those cultured in flasks. In fact, islets cultured on MG6 (n = 2) were not tested owing to complete disintegration. Islet functional quality was improved when cultured on selected biocompatible gas-permeable membranes; however, finding the best membrane requires further investigation before clinical application.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cadaver
  • Carbon Dioxide / analysis
  • Cell Count
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Culture Media, Serum-Free
  • Gases / analysis*
  • Humans
  • Insulin / metabolism*
  • Insulin Secretion
  • Islets of Langerhans / cytology*
  • Islets of Langerhans / metabolism
  • Islets of Langerhans / physiology*
  • Islets of Langerhans Transplantation
  • Membranes, Artificial*
  • Oxygen / analysis
  • Permeability
  • Tissue Donors


  • Culture Media, Serum-Free
  • Gases
  • Insulin
  • Membranes, Artificial
  • Carbon Dioxide
  • Oxygen