PDX1 gene and cytokines induce human umbilical cord mesenchymal stem cells to differentiate into islet β-like cells in vitro
10.3760/cma.j.issn.1674-1935.2011.01.015
- VernacularTitle:胰十二指肠同源框因子-1联合细胞因子诱导人脐带间充质干细胞分化为胰岛β样细胞的研究
- Author:
Bo WANG
;
Hanqing WU
;
Heshui WU
;
Chunyou WANG
- Publication Type:Journal Article
- Keywords:
Umbilical cord;
Mesenchymal stem cells;
Pancreatic and duodenal homeobox factor;
Islet β-like cells;
Cell differentiation
- From:
Chinese Journal of Pancreatology
2011;11(1):43-47
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the method how PDX1 gene modified mesenchymal stem cells (MSCs) from human umbilical cord can be differentiated into islet β-like cells in vitro. Methods Recombined adenovirus vectors inserted with PDX1 (Adxsi-CMV-PDX1) was transfected into MSCs and multiple cytokines was combined to induce differentiation. The expressions of PDX1, insulin, ngn3, glut2,NKX6.1 were examined by RT-PCR and immunofluorescence staining. The levels of insulin and C peptide secretion were examined by chemiluminescence immunoassay. Flow cytometry was used to determine the positive rate of insulin cells. Results After Adxsi-CMV-PDX1 transfection and cytokines induction, MSCs were transformed from short spindle shape to long spindle shape and aggregated into islet-like cell clusters.Dithizone staining of these cells showed bright red color. PDX1, ngn3, NKX6.1, insulin, glut 2 mRNA were expressed in cells 17d after induction. Insulin and C peptide were expressed in cytoplasm. The levels of insulin and C peptide in cell culture supernatant were (473.1 ± 51.5)mU/L and (1.61 ± 0.41)ng/ml; the levels of insulin and C peptide secretion were (964.4 ± 68.1) mU/L, (3.72 ± 1.52) ng/mL, respectively, with 25 mmol/L glucose stimulation for one hour. Insulin (+) cells rate (11.6 ± 4.8) %. Conclusions Adxsi-CMV-PDX1 combined with cytokines can induce MSCs from human umbilical cord to differentiate into islet β-like cells.They can secret insulin and C peptide, and have the sensitivity to the stimulation of glucose.
