A METHOD FOR OBTAINING MONODISPERSED CELLS FROM ISOLATED PORCINE ISLETS OF LANGERHANS

Micro or macroencapsulation of islets of Langerhans have been proposed as a bioartificial pancreas. Encapsulation of dispersed single cells instead of porcine islets should improve the oxygenation of encapsulat ed tissue. The aim of this work was, therefore, to develop techniques for dissociating porcine islets and test cell viability and function. After islet isolation and purification, islets were dispersed into sin gle cells with collagenase and DNAse in either an extracellular type i onic solution or a UW solution. After culture, islets or cells were pe rfused with Krebs buffer. Two consecutive stimulations from 2.8 mM to 20 mM glucose were performed. Viability of cells (trypan blue) was hig her than 85% after dispersion in ES or UW solutions. Islets or dispers ed cells responded similarly to both stimulations with a return to bas al rate between stimulations. No difference was found between cell fun ction cultured during 18 hours or 6 days. However, islet function was improved by a long period of culture. In conclusion, this study demons trates that dissociated cells performed as well as native islets up to six days culture.