Production of Islet-Like Structures from Neonatal Porcine Pancreatic Tissue in Suspension Bioreactors

摘要

The aim of this study was to develop a scaleable process to expand pancreatic endocrine tissue (i.e.,aggregates or islet-like structures) in suspension bioreactors.Key issues addressed included (i) serum-free media,(ii) cell inoculation density,(iii) medium pH,and (iv) aggregate dissociation.Suspension bioreactors were inoculated with pancreatic neonatal tissue and operated under controlled conditions for a 9-day period.Medium studies showed that a new serum-free medium developed in our laboratory was capable of supporting endocrine cell expansion.An inoculation density of 127,000 cells/mL resulted in more than a 7.5-fold increase in the number of insulin-positive cells after 9 days.The resulting population consisted of single cells and many islet-like aggregates that contained all of the endocrine cell types (including insulin-positive,glucagon-positive,somatostatin-positive,and pancreatic polypeptide-positive cells).Furthermore,the cell aggregates exhibited a glucose-responsive behavior.This study represents a significant milestone on the path to the effective expansion of human islet-like tissue in bioreactors that may be used for cell therapy to treat Type 1 diabetes.