1. Kerman Physiology Research Center, Kerman University of Medical Science, Kerman, Iran
Abstract
Introduction
Diabetes mellitus type 1 is a form of diabetes mellitus in which not enough insulin is produced. to this date, no definitive way to cure the disease has not been discovered. mesenchymal stem cells (mscs) are potent and self-renewable cells that can differentiate mesothermic tissues. the purpose of this study was to investigate the differentiation of human adipose tissue-derived stem cells (ascs) into insulin-producing cells (ipcs) using minimal differentiation factors in order to obtain an appropriate cellular source to treat diabetes.
Methods
Ascs were collected by liposuction aspiration and induction of differentiation was performed in two steps. in the pre-differentiation stage, a low-glucose enzyme containing nicotinamide, beta mercaptoethanol and 20% fetal bovin serum (fbs) was used in the differentiation step of the fast-glucose, nicotinamide and beta mercaptoethanol without fbs. the cell differentiation was performed by morphology, dithizone (dtz) staining, or rt-pcr (reverse transcription polymerase chain reaction). to evaluate the performance of differentiated cells, insulin secretion was measured by elisa method.
Results
According to gel electrophoresis, expression of pax4, glut2, pdx-1 and insulin genes in mrna level was observed. bata-actin was performed as an internal control. elisa data was showed significant differences in insulin secretion at day 28 compared to day 14 as well as first day. dtz-stained cellular clusters appeared after 28 days in the culture flask.
Conclusion
Human ascs can be converted to ipcs with the least differentiation factor.
