Production of islet-like structures from neonatal porcine pancreatic tissue in suspension bioreactors.
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.
Chawla
Meera
M
Pharmaceutical Production Research Facility, PPRF, Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Bodnar
Cheryl A
CA
Sen
Arindom
A
Kallos
Michael S
MS
Behie
Leo A
LA
D000818:Animals; D000831:Animals, Newborn; D019149:Bioreactors; D002478:Cells, Cultured; D003470:Culture Media; D004703:Endocrine System; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D013535:Suspensions; D013552:Swine
D003470:Culture Media; D013535:Suspensions; D005947:Glucose