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Efficient Generation of Pancreas/Duodenum Homeobox Protein 1+ Posterior Foregut/Pancreatic Progenitors from hPSCs in Adhesion Cultures
Published on: March 27, 2019
Human placenta - stem cell source for obtaining pancreatic progenitors
Sergiu Şuşman1, Dan Rus-Ciucă, Olga Soriţău
1Department of Histology, "Iuliu Hatieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania; serman_s@yahoo.com.
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.
Insights
Placental stem cells can be transformed into pancreatic progenitors using specific differentiation protocols. This offers a potential new avenue for cell therapy in treating diabetes.
Area of Science:
- Stem cell biology
- Endocrinology
- Regenerative medicine
Background:
- Type 1 diabetes results from insufficient insulin-producing beta cells.
- Cell therapy using stem cells is a promising approach for diabetes treatment.
- Understanding pancreatic development is crucial for generating functional beta cells.
Purpose of the Study:
- To investigate the potential of amniotic membrane-derived stem cells for pancreatic progenitor generation.
- To establish a differentiation protocol for obtaining pancreatic progenitors from placental stem cells.
- To evaluate the immunological properties of these cells for potential transplantation.
Main Methods:
- A three-stage differentiation protocol was applied to isolated amniotic membrane stem cells.
- Differentiation was guided by modulating glucose concentration, substrate type (collagen + laminin), nicotinamide, and exendin-4.
- Cellular analysis included immunofluorescence, gene expression (RT-PCR), and morphological assessment.
Main Results:
- Differentiated cells expressed key pancreatic progenitor transcription factors (Pdx1 and PAX4).
- Cells formed islet-like clusters and showed positive dithizone staining.
- Immunocytochemistry confirmed insulin expression in the differentiated cells.
Conclusions:
- Placental stem cells can be successfully differentiated into pancreatic progenitors.
- The established protocol provides a method for generating pancreatic progenitors from amniotic membrane-derived stem cells.
- These findings support the potential of placental stem cells in cell-based therapies for diabetes.
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