To be successful, yet, IPCs must possess physiologically appropriate regulation of insulin secretion [5, 6], including sensing circulating glucose concentrations and secreting insulin in response to physiological glucose concentrations appropriately without risk of neoplastic transformation [7, 8]. Nowadays, unresolved obstacles associated with differentiation of stem cells into IPCs include maturation of the insulin secretory pathways and mechanisms responsible for sensing ambient glucose concentrations as well as lack of sufficient development of the insulin processing
machinery [9, 10]. Atomic force microscopy (AFM) has been widely SP600125 mw used in cell biology PND-1186 solubility dmso studies, especially of both cellular and subcellular structures and topographical morphology [11, 12], because of its ability to image biological samples at nanometer resolutions. Differences in cell morphology can likely reveal the reason why there is great difference in cellular function. Thus, we compared the differences in morphology and function between normal human pancreatic beta cells and IPCs derived from human adipose-derived stem cells (hADSCs). Moreover, we examined the relationship between cell morphology and function. At the molecular level, we found that although IPCs had a similar distribution of membrane proteins to normal pancreatic beta cells, they still could not mimic the physiological regulation of insulin secretion performed by normal pancreatic
beta cells. We propose that the difference in physiological function between these two kinds this website of cells is due to the difference in the nanostructure of their cell membranes. Methods Isolation and differentiation of MSCs from human adipose Calpain tissue Human adipose tissue was obtained from four donors, two males and two females. Informed consent was obtained from participating donors according
to procedures approved by the Ethics Committee at the Chinese Academy of Medical Sciences. Experiments were performed according to the ethical standards formulated in the Helsinki Declaration. The isolated and differentiated procedure was described by Shi et al. . In order to authenticate the phenotypes of mesenchymal stem cells (MSCs), flow cytometric analysis of hADSCs was performed using antibodies for CD59, CD34, CD44, CD45, CD105, CD13, and HLA-DR (BD Biosciences, Franklin Lakes, NJ, USA). Culture of normal human pancreatic beta cells Normal human pancreatic beta cells were obtained commercially (HUM-CELL-0058, Wuhan Pricells Biotechnology & Medicine Co., Ltd., Wuhan, China). Expansion medium contained MED-0001 and 5 ng/mL rhEGF, 5 μg/mL rhinsulin, 5 μg/mL transferrin, 10 nM T3, 1.0 μM epinephrine, 5 μg/mL hydrocortisone, 10% fetal bovine serum (all expansion media were from Wuhan Pricells Biotechnology & Medicine Co., Ltd.). The cells were cultured in complete medium in T-25 tissue culture flasks that have been coated with collagenase at 37°C in 5% CO2.