Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro
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Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro
Journal of Chongqing Medical UniversityVol. 38, Issue 11, Pages: 1617-1621(2014)
作者机构:
1. 重庆医科大学附属儿童医院发育疾病研究教育部重点实验室,重庆,400014
2. 重庆医科大学附属儿童医院分子生物中心,重庆,400014
3. 重庆医科大学附属儿童医院呼吸中心,重庆,400014
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Published:2014,
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Gao Li, Zhu Hua, Zhang Rong, et al. Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro. [J]. Journal of Chongqing Medical University 38(11):1617-1621(2014)
DOI:
Gao Li, Zhu Hua, Zhang Rong, et al. Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro. [J]. Journal of Chongqing Medical University 38(11):1617-1621(2014)DOI:
Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro
Objective:To explore the method of using 5-bromo-2-deoxyuridine(BrdU) to label human umbilical cord mesenchymal stem cells(hUC-MSCs),and to explore its effect on cell proliferation and apoptosis. Methods:HUC-MSCs were isolated and cultured by direct adherent method,then the surface marker and potential ability of multi-direction differentiation were analyzed .The fifth passage of cells were labeled with BrdU at concentrations of 10,15,20,25 ?滋mol/L,excluding BrdU as control group. Cells were culti-vated respectively for 24,48,72,96 h. Trypan blue testing was used to detect cell vitality,MTT method was used to analyze cell pro-liferation and AnnexinV-FITC assay was used to detect cell survival rate and apoptosis rate. Results:Trypan blue testing showed that the living cells was more than 96% and there was no statistical significance between experimental group and control group(compar-ison among different concentration groups,P=0.89;comparison among different time points,P=0.61;interaction between time and dif-ferent concentrations,P=0.89). MTT method detected that there was no significant difference in proliferation activity between labeled cells and unlabeled cells(P >0.05). There were differences between cell proliferation and apoptosis rate(proliferation rate,F=12.02,P<0.001;apoptosis rate,F=6.148,P=0.009). Conclusion:The tracing method of BrdU labeling hUC-MSCs is safe and ef-fective. The best labeling concentration and time is 20 μmol/L and 72 h.
关键词
人脐带间充质干细胞5-溴脱氧尿嘧啶核苷增殖凋亡
Keywords
human umbilical cord mesenchymal stem cells5-bromo-2-deoxyuridineproliferationapoptosis