Abstract | OBJECTIVE: METHODS: The iron overload model of MSC was established by in vitro addition of ferric ammonium citrate (FAC) into culture medium. Cell proliferation and apoptosis were determined by Annexin V/PI double staining and population doubling time (DT) respectively. Co-culture system was used to assess the hematopoietic support capacity of UC-MSC in different groups. Thereafter the ROS level was detected with fluorescent probe 2', 7'-dichlorofluorescin diacetate ( DCFH-DA). And the ROS related signaling factors of p-p38MAPK, p38 MAPK, P53 were measured by Western blot. RESULTS: (1) The DT of UC-MSC in iron overload group was significantly longer than that of control ((24.43 ± 2.72) h vs (16.03 ± 2.31) h, P < 0.05). But the difference was insignificant after two passages (P > 0.05). (2) Apoptosis in iron overload group was higher than that of control (12.75% ± 0.32% vs 3.63% ± 0.80%, P < 0.05). (3) The colony forming capacity of mononuclear cell (MNC) co-cultured with UC-MSC of iron overload group for 1/2 weeks significantly decreased. (4) The ROS level of UC-MSC with iron overload was higher than that of control in time and concentration-dependent fashions and it peaked at 400 µmol/L of FAC for 12 h (1499 ± 86 vs 548 ± 97, P < 0.05). (5) The expressions of p-p38MAPK and P53 increased in response to FAC compared with control. But such an effect was partially inhibited after the use of antioxidants. CONCLUSIONS:
Iron overload may impair the proliferation, survival and hematopoiesis supportive function of UC-MSC by enhancing the generation of ROS. And ROS stimulates the signaling pathways of p-p38MAPK and P53.
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Authors | Wen-yi Lu, Ming-feng Zhao, Xiao Chai, Juan-xia Meng, Nan Zhao, Sajin Rajbhandary, Xin-nü Xu, Li Ma, Yu-ming Li |
Journal | Zhonghua yi xue za zhi
(Zhonghua Yi Xue Za Zhi)
Vol. 93
Issue 12
Pg. 930-4
(Mar 26 2013)
ISSN: 0376-2491 [Print] China |
PMID | 23863680
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Culture Media
- Reactive Oxygen Species
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Topics |
- Cell Proliferation
- Cells, Cultured
- Culture Media
- Hematopoietic System
- Humans
- Iron Overload
- Mesenchymal Stem Cells
(cytology)
- Reactive Oxygen Species
(metabolism)
- Signal Transduction
- Umbilical Cord
(cytology)
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