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Life Sci ; 267: 118451, 2021 Feb 15.
Article in English | MEDLINE | ID: mdl-32956667

ABSTRACT

AIMS: We investigated the in vitro differentiation of adult rat PDESCs into ß-like cells through supplementation of different combinations of GABA, BMP7, and Activin A in basic culture media. MATERIALS AND METHODS: The PDESCs were cultured using different inducement combinations for 28 days and microscopy, dithizone (DTZ) staining, immunohistochemical staining, real-time PCR, and glucose-stimulated insulin secretion (GSIS) assay were used to delineate the differentiation inducement potential of these combinations. KEY FINDINGS: The results show that after 28 days, the PDESCs were differentiated into ICCs containing insulin-secreting ß-like cells in different groups treated with A + B, A + G, B + G, and A + B + G but not in the control group. Upon DTZ staining the cells in ICCs were stained crimson red, demonstrating the presence of ß-like cells in ICCs and the immunohistochemistry showed the expression of Pdx1 and insulin in ICCs. Further, on 28 d the expression of Pdx1 and insulin mRNA was high in inducement groups as compared to the control group and ß-like cells in ICCs also secreted insulin and C-peptide upon glucose stimulation. Thus, the supplementation of GABA, BMP7, and Activin A in different combinations in basic culture media can induce the in vitro differentiation of PDESCs into ICCs containing ß-like cells. SIGNIFICANCE: The in vitro development of ß-like cells is a herald for cell therapy of diabetic patients and our results are a step closer towards finding the cure for diabetes.


Subject(s)
Cell Differentiation/drug effects , Insulin-Secreting Cells/metabolism , Stem Cells/metabolism , Activins/metabolism , Activins/pharmacology , Animals , Bone Morphogenetic Protein 7/metabolism , Bone Morphogenetic Protein 7/pharmacology , Cell Culture Techniques/methods , Cell Line , Cells, Cultured , Culture Media/chemistry , Insulin/metabolism , Insulin Secretion , Pancreatic Ducts/cytology , Pancreatic Ducts/metabolism , Rats , Rats, Sprague-Dawley , Stem Cells/cytology , gamma-Aminobutyric Acid/metabolism , gamma-Aminobutyric Acid/pharmacology
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