A simple method to induce differentiation of murine bone marrow mesenchymal cells to insulin-producing cells using conophylline and betacellulin-delta4.
The present study was conducted to establish a method to induce differentiation of bone marrow (MB)-derived mesenchymal cells into insulin-producing cells. When mouse BM-derived mesenchymal cells were cultured for 60 days in medium containing 10% fetal calf serum and 25 mM glucose, they expressed insulin. Addition of activin A and betacellulin (BTC) accelerated differentiation, and immunoreactive insulin was detected 14 days after the treatment. Insulin-containing secretory granules were observed in differentiated cells by electron microscopy. Treatment of BM-derived mesenchymal cells with conophylline (CnP) and BTC-delta4 further accelerated differentiation, and mRNA for insulin was detected 5 to 7 days after the treatment. Mesencymal cells treated with CnP and BTC-delta4 responded to a high concentration of glucose and secreted mature insulin. When these cells were transplanted into streptozotocin-treated mice, they markedly reduced the plasma glucose concentration, and the effect continued for at least 4 weeks. These results indicate an efficacy of the combination of CnP and BTC-delta4 in inducing differentiation of BM-derived mesenchymal cells into insulin-producing cells.
Hisanaga
Etsuko
E
Institute for Molecular & Cellular Regulation, Gunma University, Maebashi, Japan.
Park
Kee-Yong
KY
Yamada
Satoko
S
Hashimoto
Hiromi
H
Takeuchi
Toshiyuki
T
Mori
Masatomo
M
Seno
Masaharu
M
Umezawa
Kazuo
K
Takei
Izumi
I
Kojima
Itaru
I
D017667:Adipocytes / Q000187:drug effects / Q000502:physiology; D000818:Animals; D066259:Betacellulin; D001786:Blood Glucose / Q000032:analysis; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000502:physiology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D007328:Insulin / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D014748:Vinca Alkaloids / Q000494:pharmacology*
D066259:Betacellulin; D001786:Blood Glucose; C587852:Btc protein, mouse; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D014748:Vinca Alkaloids; C099167:conophylline