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Background and purpose: Regarding the effective role of plant flavonoids in the prevention and treatment of diabetes mellitus, in this investigation, induction of P19 stem cells differentiation into insulin producing cells by flavonoids of methanol extract of Cichorium intybus L. leaf was examined. Materials and methods: Initially, flavonoids of methanol extract of Cichorium intybus L. leaf were concentrated. The P19-derived embryonic bodies were cultured in α-MEM (Minimum Essential Medium) containing 15% fetal bovine serum for three days. To induce the differentiation, the cells treated with the concentrations of 50, 100 and 200 µg/ml of flavonoids of methanol extract of Cichorium intybus L. leaf for 12 days. Untreated P19 cells and embryonic bodies were used as controls. Dithizone staining, immunoflorecence and real-time polymerase chain reaction (RT-PCR) techniques were used to demonstrate the differentiated cells. Results: Differentiated P19 cells by different concentrations of extract shown positive reaction to dithizone staining. Most percentage of positive dithizone cells was reached in concentration of 100 µg/ml. Immunoflorecence method showed that differentiated P19 cells were able to express the pancreas beta cell-specific markers. Pdx-1 gene expression in the cells was demonstrated by RT-PCR technique. Conclusion: The flavonoids of methanol extract of Cichorium intybus L. leaf are able to induce differentiation of P19 stem cells to insulin producing cells.

Keywords: Methanol extract, flavonoid, Cichorium intybus L. leaf, P19 cells, insulin-producing cells, Pdx-1 gene, pancreas beta cells

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