ABSTRACT
Objective: Worldwide, diabetes mellitus [DM] is an ever-increasing metabolic disorder
A promising approach to the treatment of DM is the implantation of insulin producing cells [IPC] that have been derived from various stem cells. Culture conditions play a pivotal role in the quality and quantity of the differentiated cells. In this experimental study, we have applied various culture conditions to differentiate human umbilical cord matrix-derived mesenchymal cells [hUCMs] into IPCs and measured insulin production
Materials and Methods: In this experimental study, we exposed hUCMs cells to pancreatic medium and differentiated them into IPCs in monolayer and suspension cultures
Pancreatic medium consisted of serum-free Dulbecco's modified eagle's medium Nutrient mixture F12 [DMEM/F12] medium with 17.5 mM glucose supplemented by 10 mM nicotinamide, 10 nM exendin-4, 10 nM pentagastrin, 100 pM hepatocyte growth factor, and B-27 serum-free supplement. After differentiation, insulin content was analyzed by gene expression, immunocytochemistry [IHC] and the chemiluminesence immunoassay [CLIA]
Results: Reverse transcription-polymerase chain reaction [RT-PCR] showed efficient expressions of NKX2.2, PDX1 and INSULIN genes in both groups. IHC analysis showed higher expression of insulin protein in the hanging drop group, and CLIA revealed a significant higher insulin production in hanging drops compared with the monolayer group following the glucose challenge test
Conclusion: We showed by this novel, simple technique that the suspension culture played an important role in differentiation of hUCMs into IPC. This culture was more ef- ficient than the conventional culture method commonly used in IPC differentiation and cultivation