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Tang X.,Central Hospital of Liaohe Oil Field | Wang Y.,Central Hospital of Liaohe Oil Field | Yi H.-B.,Second Workers Hospital of Liaohe Oil Field | Pang T.-S.,Central Hospital of Liaohe Oil Field
Chinese Journal of Tissue Engineering Research | Year: 2013

BACKGROUND: Intramyocardial transplantation of autologous umbilical cord-derived mesenchymal stem cells for repair of myocardial tissue damage is paid increasing attention in the cardiovascular field. OBJECTIVE: Human umbilical cord mesenchymal stem cells were isolated and cultured. Passage 2 human umbilical cord mesenchymal stem cells were treated with various concentratins of 5-azacytidine (2.5, 5, 10, 20, 40, 80 μmol/L) for 24 hours. After removal of 5-azacytidine, cells were cultured for another 4 weeks. RESULTS AND CONCLUSION: Before 5-azacytidine treatment, filament-like structures or particles were not observed in the cells, but the amount of cytoplasm was less and uniform, nuclear/cytoplasm ratio was high, cells exhibited typical fusiform appearance and grew in a swirl-like manner, and nucleolus was obvious. After treatment with 5-azacytidine for 24 hours, some cells died in each group, and typical fusiform appearance turned into stick-like or column-like appearance, especially in the 40 and 80 μmol/L 5-azacytidine groups. Reverse transcription-PCR results showed that atrial natriuretic peptide and α-skeletal actin gene expression levels were detected on human umbilical cord mesenchymal stem cells after treatment with 2.5 or 40 μmol/L 5-azacytidine for 4 weeks or with 5, 10, 20 μmol/L 5-azacytidine for 1, 2, 3 and 4 weeks. These findings suggest that 5-azacytidine-induced human umbilical cord mesenchymal stem cells express the specific gene of myocardiocytes. Source


Tang X.,Central Hospital of Liaohe Oil Field | Wang N.-N.,Central Hospital of Liaohe Oil Field | Yi H.-B.,Second Workers Hospital of Liaohe Oil Field | Wang Y.,Central Hospital of Liaohe Oil Field | Pang T.-S.,Central Hospital of Liaohe Oil Field
Chinese Journal of Tissue Engineering Research | Year: 2013

Background: Human umbilical cord mesenchymal stem cells possess multi-differentiation potential. There are few studies describing whether human umbilical cord mesenchymal stem cells can be induced to differentiate into cardiomyocytes. Objective: To investigate cardiac troponin I expression in human umbilical cord mesenchymal stem cells after 5-azacytidine induction in vitro. Methods: Passage 2 human umbilical cord mesenchymal stem cells were digested, centrifuged and deposited. Cell suspension was prepared and cell concentration was adjusted to be 2×107/L. Then cells were cultured for 24 hours after addition of various concentrations of 5-azacytidine (80, 40, 20, 10, 5, 2.5, 0 μmol/L). After removal of 5-azacytidine, cells were cultured for another 4 weeks. Results and Conclusion: After treatment with 5-azacytidine for 24 hours, some cells died in each group, and typical fusiform appearance turned into stick-like or column-like appearance, especially in the 40 and 80 μmol/L 5-azacytidine groups. Immunohistochemical staining results showed that cardiac troponin I expression was detected in human umbilical cord mesenchymal stem cells in 5, 10, 20, 40, 80 μmol/L 5-azacytidine groups after induction for 2 weeks, but it was not detected in the control and 2.5 μmol/L 5-azacytidine groups. Five high-fold fields were selected for cell counting. The number of cardiomyocytes was not significantly different between 80, 40, 20, 10 μmol/L 5-azacytidine groups, but it was significantly higher compared to that in the 5 μmol/L 5-azacytidine group. After 5-azacytidine treatment for 4 weeks, typical sarcomeres and cardiac atrium particles were observed in some human umbilical cord mesenchymal stem cells. Such changes were not observed in the control group in which 5-azacytidine induction was not used. These findings suggest that human umbilical cord mesenchymal stem cells were induced to cardiomyocyte-like cells by 5-azacytidine. Source

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