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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


Tang X.,The Central Hospital of Liaohe Oil Field | Wang Y.,The Central Hospital of Liaohe Oil Field | Yi H.-b.,Second Workers Hospital of Liaohe Oil Field | Pang T.-s.,The Central Hospital of Liaohe Oil Field
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2011

BACKGROUND: Studies addressing the induced effect of different-concentration 5-azacytidine on the in vitro differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cardiomyocytes are fewer. OBJECTIVE: To select a suitable concentration of 5-azacytidine to induce differentiatiown of hUC-MSCs into cardiomyocytes in vitro. METHODS: Second passage hUC-MSCs were cultured in culture medium containing different -concentration 5-azacytidine (2.5, 5, 10, 20, 40, 80 μmol/L, respectively). After incubating for 24 hours, the medium was changed to complete medium without 5-azacytidine. Thereafter, the cells were cultivated for 4 weeks. Two weeks after the cells were treated with 5-azacytidine, cardiac troponin I was assayed by immunohistochemical methods and the ratio of positive cells for cardiac troponin I was counted. RESULTS AND CONCLUSION: Seven days after hUC-MSCs were treated with 5-azacytidine, the cellular morphology began to change. The cells arranged closely and in parallel. The cellular morphology of 40 and 80 μmol/L groups changed obviously. At 4 weeks, the refraction and activity of cultured cells declined in the 40 and 80 μmol/L groups, and some of the cells died. At 2 weeks, cardiac troponin I was positive for hUC-MSCs in 5, 10, 20, 40 and 80 μmol/L groups and negative in 2.5 μmol/L and control groups. The ratio of positive cells for cardiac troponin I in 10, 20, 40 and 80 μmol/L groups was significantly higher than that of 5 μmol/L group (P < 0.05). l0 μmol/L 5-azacytidine is the optimal concentration for hUC-MSCs differentiation into cardiomyoctyes. Source

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