Promotion of beta-cell differentiation by the alkaloid conophylline in porcine pancreatic endocrine cells.
We previously found that conophylline, an alkaloid isolated from the leaves of Ervatamia microphylla, induced beta-cell differentiation in rat pancreatic acinar carcinoma cells and in cultured fetal rat pancreatic tissue and that it also decreased the blood glucose level in streptozotocin-treated fetal rats. In the present research, we looked into the effect of conophylline on the differentiation of newborn pig pancreatic endocrine cells into insulin-secreting cells. Conophylline potentiated the differentiation of monolayer cells into insulin-producing cells in the presence of nicotinamide in 3 weeks. Next we prepared islet-like cell clusters (ICC). Cononophylline together with nicotinamide also increased the number of insulin-producing cells and the insulin content in ICC in 3-6 weeks. The ICC thus prepared were sensitive to the glucose concentration for the insulin secretion. Conophylline increased the mRNA expression of PDX-1, neurogenin3, neuroD/Beta2, and insulin in ICC. Thus, the vinca alkaloid conophylline potentiated beta-cell differentiation in porcine pancreatic endocrine-rich cells in cluster cultures. Pig pancreatic cells are practical candidate for use in transplantation therapy. Conophylline may thus be useful for the large-scale preparation of porcine insulin-producing cells for the regeneration therapy of type-1 diabetes mellitus.
Kawakami
M
M
Center for Chemical Biology, School of Fundamental Science and Technology, Keio University, Yokohama, Japan.
Hirayama
A
A
Tsuchiya
K
K
Ohgawara
H
H
Nakamura
M
M
Umezawa
K
K
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism; D015394:Molecular Structure; D009536:Niacinamide / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D034421:Sus scrofa; D013552:Swine; D014748:Vinca Alkaloids / Q000737:chemistry / Q000494:pharmacology*
D015415:Biomarkers; D007328:Insulin; D014748:Vinca Alkaloids; C099167:conophylline; D009536:Niacinamide; D005947:Glucose