Characterization of human islet-like structures generated from pancreatic precursor cells in culture.
This study addresses the characterization of human islet-like structures generated from a newly discovered sparse population of precursor cells (Petropavlovskaia and Rosenberg, 2002) in the human pancreas. These cells may be progenitor cells capable of producing pancreatic cells suitable for the treatment of type 1 diabetes. The cells were cultured successfully in non-adherent stationary cultures and yielded, as an important first step, a 1.9-fold expansion in a serum-free medium developed specifically for this cell type. This expanded population grew as pancreatic cell aggregates, which were analyzed for islet-like characteristics. Specifically, through RT-PCR analyses and functionality assays, we show that cells within the population expressed all four of the endocrine hormone genes and proteins (insulin, glucagon, somatostatin, pancreatic polypeptide). As well, the expanded pancreatic precursor cell population exhibited glucose responsiveness although the produced cells appeared to be still primitive in nature.
Bodnar
Cheryl A
CA
Pharmaceutical Production Research Facility (PPRF), SCHULICH School of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
Sen
Arindom
A
Kallos
Michael S
MS
Behie
Leo A
LA
Petropavlovskaia
Maria
M
Rosenberg
Lawrence
L
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D002096:C-Peptide / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D005934:Glucagon / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010191:Pancreatic Polypeptide / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013004:Somatostatin / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D046509:Tissue Culture Techniques / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics
D002096:C-Peptide; D003470:Culture Media; D016895:Culture Media, Serum-Free; D018398:Homeodomain Proteins; D007328:Insulin; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D013004:Somatostatin; D010191:Pancreatic Polypeptide; D005934:Glucagon; D005947:Glucose; D015056:1-Methyl-3-isobutylxanthine