A Simple Method to Induce Differentiation of Murine Bone Marrow Mesenchymal Cells to Insulin-producing Cells Using Conophylline and Betacellulin-delta4

Abstract

Abstract. 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 BMderived 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.

Betacellulin Overexpression in Mesenchymal Stem Cells Induces Insulin Secretion In Vitro and Ameliorates Streptozotocin-Induced Hyperglycemia in Rats

“…Our cells presented all the characteristics required, as stated by the International Society for Cellular Therapy [27], confirming MSC identity and plasticity. On the other hand, in agreement with our results, Hisanaga et al have reported the in vitro differentiation of murine MSCs by the addition of BTC to the medium [44].…”

Cell Replacement Strategies Aimed at Reconstitution of the β-Cell Compartment in Type 1 Diabetes

“…In a recent review of the subject, Domínguez-Bendala et al (65) echoed the growing perception that these mesodermal cells are intrinsically incapable of becoming bona fide endodermal b-cells. Their directed differentiation and subsequent transplantation has met only with partial success in preclinical models (66)(67)(68)(69)(70)(71)(72)(73)(74)(75). However, when directly infused in an undifferentiated state onto diabetic subjects, a variable degree of endogenous regeneration has been observed (76)(77)(78)(79).…”

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

“…MSCs were isolated from many organs and tissues, such as bone marrow, fat tissue, umbilical cord blood, pancreas etc. (Hisanaga et al 2008;Sun et al 2007;Zhang et al 2009b). Mesenchymal stem cells are a heterogeneous population of cells capable to multidirectional differentiation (Ding et al 2011).…”

“…A simple method to induce differentiation of murine bone marrow MSCs to insulin-producing cells was presented by Hisanaga et al (2008). To induce bone marrow mesenchymal stem cells (BM-MSCs) differentiation into pancreatic endocrine cells, the cells were cultured in the basic medium in the presence of activin-A, conophylline and betacellulin-d4 (Table 3).…”

“…Differentiated cells secreted mature insulin and responded to glucose stimulation in vitro. When these cells were transplanted to diabetic mice, they markedly reduced the glucose concentration (Hisanaga et al 2008). Sun et al (2007) reported that BMMSCs can be differentiated into IPCs by a three-stage protocol.…”