Human placenta - stem cell source for obtaining pancreatic progenitors.

OBJECTIVES: The apparition of sugar diabetes is produced by the decrease of the number and capacity of beta cells to secrete insulin. Cell mass recovery through cell therapy might be one of the solutions for treating this disease. The use of various cell sources of different differentiation grades has been tried over the last years. Decoding the molecular mechanisms of the pancreatic morphogenesis is essential for obtaining cells having a phenotype, which would be very similar to the mature cells located in the pancreatic endocrine component. In this study, in order to obtain pancreatic progenitors, we used stem cells harvested from the mesenchymal component of the amniotic membrane, cells with particular immunological properties, which are effective in transplant.

MATERIALS AND METHODS: Isolated cells from the placenta (amniotic membrane) have undergone a three-stage differentiation protocol. The modulation of glucose concentration, the type of substrate (collagen + laminin) and the use of nicotinamide and exedin-4 were the main selective conditions of differentiation microenvironment. The differentiated cells were analyzed from the point of view of proteins (immunofluorescence - IF), gene expression (real-time polymerase chain reaction - RT-PCR) and morphological changes.

RESULTS: Isolated cells from the placenta membrane induced for pancreatic differentiation expressed transcription factors, which are characteristic for pancreatic progenitors (Pdx1 and PAX4). During the experiment, the cells modified their morphology by forming islet-like clusters. They were positively for dithizone staining and expressed insulin as shown by immunocytochemistry.

CONCLUSIONS: The isolated cells from the placenta can be differentiated towards pancreatic progenitors by using specific protocols.