MafA-positive insulin-producing cells derived from rat adipose stem cells are efficient in glucose hemostasis in vitro

Dian Dayer; Elham  Ghalami Negad; Seyed Javad Hosseni

doi:10.53730/ijhs.v6nS7.13403

Authors

Dian Dayer PhD of Molecular Medicine, Cellular and Molecular Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Elham Ghalami Negad MSC of Cellular and Molecular Sciences, Group of Biotechnology, Institute of Persian Gulf, Persian Gulf University, Bushehr 75169, Iran
Seyed Javad Hosseni
sjh1348@gmail.com
PhD in Cellular and Molecular Biology, Group of Biotechnology, Institute of Persian Gulf, Persian Gulf University, Bushehr 75169, Iran and Department of Biological Science and Technology, Faculty of Nano and Bio Science and Technology, Persian Gulf University, Bushehr 75169, Iran

Keywords:

Insulin-producing cells, Adipose tissue-derived mesenchymal stem cells, Glut2, ENO1, IGF-I receptor, Glucagon

Abstract

Objective: MafA has been recognized as a specific pancreatic β-cells activator for Insulin expression. This study investigated the changes in glucose transport and consumption by MafA-positive Insulin-producing cells resulting from the differentiation of rat adipose tissue-derived mesenchymal stem cells. Methods: Adipose tissue-derived mesenchymal stem cells were differentiated into Insulin-producing cells in a 14-day protocol using nicotinamide and ITS. The ability to produce Insulin was confirmed by adipogenesis differentiation capacity and expression of Oct4, Nanog, and Sox2. Insulin and Glucagon concentrations in Insulin-producing cells were determined by ELISA. The ability of Insulin-producing cells for the presence of MafA, Glut2, ENO1, and IGF-I receptor genes was determined by Western blotting analysis. The amounts of Glucokinase and Pyruvate Carboxylase activities were measured by Fluorometric and Colorimetric methods, respectively. Results: The un-differentiated cells showed a promising capacity to differentiate toward adipocytes. The adipose tissue mesenchymal stem cells revealed significantly elevated levels of Oct4, Nanog, and Sox2 compared to differentiated adipocytes. The cells that differentiated into Insulin-producing cells secreted considerably more Insulin and Glucagon than undifferentiated cells.

Downloads

Download data is not yet available.

References

Objective: MafA has been recognized as a specific pancreatic β-cells activator for Insulin expression. This study investigated the changes in glucose transport and consumption by MafA-positive Insulin-producing cells resulting from the differentiation of rat adipose tissue-derived mesenchymal stem cells. Methods: Adipose tissue-derived mesenchymal stem cells were differentiated into Insulin-producing cells in a 14-day protocol using nicotinamide and ITS. The ability to produce Insulin was confirmed by adipogenesis differentiation capacity and expression of Oct4, Nanog, and Sox2. Insulin and Glucagon concentrations in Insulin-producing cells were determined by ELISA. The ability of Insulin-producing cells for the presence of MafA, Glut2, ENO1, and IGF-I receptor genes was determined by Western blotting analysis. The amounts of Glucokinase and Pyruvate Carboxylase activities were measured by Fluorometric and Colorimetric methods, respectively. Results: The un-differentiated cells showed a promising capacity to differentiate toward adipocytes. The adipose tissue mesenchymal stem cells revealed significantly elevated levels of Oct4, Nanog, and Sox2 compared to differentiated adipocytes. The cells that differentiated into Insulin-producing cells secreted considerably more Insulin and Glucagon than undifferentiated cells. The differentiated cells showed significantly higher amounts of MafA, Glut2 and IGF-I receptor protein expression compared with undifferentiated cells. While At the same time, the expression of ENO1 protein was not significantly different between the two groups. Conclusion: The obtained Insulin-producing cells seem to exhibit the ability to absorb glucose naturally, perform glycolysis, metabolism-secretion coupling and glucose-dependent Insulin secretion in vitro.