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 encapsulated 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 single cells with collagenase and DNAse in either an extracellular type ionic solution or a UW solution. After culture, islets or cells were perfused with Krebs buffer. Two consecutive stimulations from 2.8 mM to 20 mM glucose were performed. Viability of cells (trypan blue) was higher than 85% after dispersion in ES or UW solutions. Islets or dispersed cells responded similarly to both stimulations with a return to basal rate between stimulations. No difference was found between cell function cultured during 18 hours or 6 days. However, islet function was improved by a long period of culture. In conclusion, this study demonstrates that dissociated cells performed as well as native islets up to six days culture.
Pueyo
M E
ME
INSERM U341, Service de Diabétologie, Hôtel-Dieu, Paris, France.
Darquy
S
S
Arbet-Engels
C
C
Poitout
V
V
Di Maria
S
S
Gangnerau
M N
MN
Reach
G
G
D000704:Analysis of Variance; D000818:Animals; D001187:Artificial Organs; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D017364:Collagenases / Q000737:chemistry; D004339:Drug Compounding; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010101:Oxygen Consumption / Q000502:physiology; D013552:Swine
D007328:Insulin; D017364:Collagenases; D005947:Glucose