Long-Term Function of Porcine Islets and Single Cells Embedded in Barium-Alginate Matrix

 

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Summary

The influence of alginate-embedding on the maintenance of functioning and morphological integrity in long-term culture of isolated porcine islets and islet cells was studied. Free-floating islets and islet cells served for control. Function was tested after the 1st, 2nd and 4th week. Basal and glucose-stimulated insulin secretion of embedded islets decreased slightly, but significantly after the first week (from 4.39±0.64 to 2.87±10.47 at normal and from 11.96±1.44 to 4.76±0.78 μU pro 24 h pro islet at elevated glucose concentration, p<0.05 and <0.01, resp.) and remained unchanged thereafter. Glucose-stimulation resulted in significant increases in insulin secretion at all three testings (p<0.001, <0.01 and <0.01). Single cells in alginate matrix had evenmore stable insulin secretion throughout the whole cultivation with significant increases to glucose challenge (p<0.01, <0.01 and <0.05). In contrast, insulin secretion of free-floating islet cells decreased from 5.70±1.19 to 2.04±0.64 and to 1.05±0.33 at basal conditions (p<0.01 and <0.05) and from 11.39±1.87 to 2.76±0.76 and to 2.15±0.71 μU pro 24 h pro islet under stimulation (p<0.01 and not sign.). In addition, the secretory response to glucose challenge was significant only at the first testing (p<0.05). Non-embedded islets could be tested only at the first week since after this time they dissociated to single cells. Embedded islets and single cells showed intact morphology after four weeks with trypan blue (TPB) positivity of less than 5%. The proportion of TBP positive free islet cells was <5% at start but increased to 30% by the end of the culture period. Accordingly, alginate matrix proved to be a suitable environment for isolated porcine islets and single cells to maintain their function and morphological integrity.