中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (32): 5167-5171.doi: 10.3969/j.issn.2095-4344.2015.32.015

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

以EdU体外标记人脐带间充质干细胞：5，10 µmol/L是其最适浓度

王  武，张  飞，李贵才，齐宝军，武忠炎，曹  鹏   

  1. 新疆医科大学第五附属医院骨科，新疆维吾尔自治区乌鲁木齐市  830001
  • 出版日期:2015-08-06 发布日期:2015-08-06
  • 通讯作者: 武忠炎，主任医师，副教授，硕士生导师，新疆医科大学第五附属医院骨科，新疆维吾尔自治区乌鲁木齐市830001
  • 作者简介:王武，男，1982年生，陕西省西安市人，汉族，2010年新疆医科大学毕业，硕士，主治医师，主要从事创伤骨科研究。 并列第一作者：张飞，男，1988年生，陕西省延安市人，汉族，2015年新疆医科大学毕业，硕士，医师，主要从事创伤骨科研究
  • 基金资助:

    新疆医科大学科研创新基金项目(XYDCX2014167)

EdU-labeled human umbilical cord mesenchymal stem cells in vitro: 5 and 10 µmol/L are the optimal concentrations of EdU 

Wang Wu, Zhang Fei, Li Gui-cai, Qi Bao-jun, Wu Zhong-yan, Cao Peng   

  1. Department of Orthopedic Surgery, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, Xinjiang Uygur Autonomous Region, China
  • Online:2015-08-06 Published:2015-08-06
  • Contact: Wu Zhong-yan, Chief physician, Associate professor, Master’s supervisor, Department of Orthopedic Surgery, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, Xinjiang Uygur Autonomous Region, China
  • About author:Wang Wu, Master, Attending physician, Department of Orthopedic Surgery, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, Xinjiang Uygur Autonomous Region, China Zhang Fei, Master, Physician, Department of Orthopedic Surgery, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830001, Xinjiang Uygur Autonomous Region, China Wang Wu and Zhang Fei contributed equally to this work.
  • Supported by:

    the Medical Research and Innovation Fund of Xinjiang Medical University, No. XYDCX2014167 

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摘要：

背景：EdU为新一代细胞核标记物，目前对此标记的研究较少。
目的：采取EdU标记人脐带间充质干细胞，筛选出EdU标记人脐带间充质干细胞的最适浓度。
方法：采用组织块法分离、纯化及传代培养人脐带间充质干细胞，倒置显微镜观察其形态及生长情况，流式细胞仪鉴定细胞表面标志物，并进行成脂诱导分化能力鉴定。分别采用5，10，20，50，100 µmol/L浓度的EdU标记脐带间充质干细胞24 h，选择标记率最高的优化浓度，绘制细胞增殖曲线。
结果与结论：倒置显微镜下观察细胞为贴壁生长，形态为长梭形，且EdU标记后的细胞形态与其一致，流式细胞仪检测CD44呈阳性表达，成脂诱导后具有向脂肪细胞分化的能力。5，10 µmol/L EdU的标记率最高，两种浓度标记的人脐带间充质干细胞生长曲线未见明显差异，故5，10 µmol/L是体外标记人脐带间充质干细胞的最适浓度。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 干细胞, 脐带脐血干细胞, 细胞示踪, 脐带间充质干细胞, EdU, 组织块法

Abstract:

BACKGROUND: EdU is a new nuclear marker, and currently, it is rarely reported.
OBJECTIVE: To determine the optimal concentration of EdU to label human umbilical cord mesenchymal stem cells.
METHODS: Human umbilical cord mesenchymal stem cells were isolated, purified and subcultured. Cell morphology and growth were observed under inverted microscope. Flow cytometry was used to identify cell surface markers, as well as adipogenic identification. EdU at concentrations of 5, 10, 20, 50, 100 µmol/L was used to label human umbilical cord mesenchymal stem cells for 24 hours. The optimal concentration that resulted in the highest labeling efficiency was selected, and then cell proliferation curve was drawn.
RESULTS AND CONCLUSION: Under the inverted microscope, cells grew adherently in a long spindle shape, and EdU-labeled cells had the same morphology. Flow cytometry showed that cells were positive for CD44, and had adipogenic differentiation ability. When the concentration of EdU was 5 and 10 µmol/L, the labeling efficiency was the highest, indicating that 5 and 10 µmol/L are the optimal concentrations of EdU to label human umbilical cord mesenchymal stem cells. 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cells, Pyrimidine Nucleosides

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