中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 4970-4974.doi: 10.3969/j.issn.1673-8225.2010.27.007

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

兔骨髓间充质干细胞体外BrdU标记最佳时间和标记浓度的筛选

朱  峰1，郭光华2，詹剑华2，谢  安3   

  1. 南昌大学第一附属医院，1重症医学科，2烧伤科，  3泌尿外科研究所，江西省南昌市   330006
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 通讯作者: 郭光华，主任医师，博士生导师，南昌大学第一附属医院烧伤中心，江西省南昌市 330006 guogh2000@hotmail.com
  • 作者简介:朱峰☆，男，1977年生，江西省南昌市人，汉族，南昌大学在读博士，讲师，主治医师，主要从事吸入性损伤及干细胞研究。zhujunchi@tom.com
  • 基金资助:

    江西省教育厅青年科学基金项目(GJJ09432),课题名称：“骨髓间充质干细胞移植对吸入性损伤生物学作用的研究”，

Screening of optimal labeling time and labeling concentration of rabbit bone marrow mesenchymal stem cells by 5-bromodeoxyuridine in vitro

Zhu Feng1, Guo Guang-hua2, Zhan Jian-hua2, Xie An3   

  1. 1 Department of Critical Care Medicine, 2 Department of Burn, 3Urology Institute, First Affiliated Hospital, Nanchang University, Nanchang  330006, Jiangxi Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Guo Guang-hua, Chief physician, Doctoral supervisor, Department of Burn, First Affiliated Hospital, Nanchang University, Nanchang 330006, Jiangxi Province, China guogh2000@hotmail.com
  • About author:Zhu Feng☆, Studying for doctorate, Lecturer, Attending physician, Department of Critical Care Medicine, First Affiliated Hospital, Nanchang University, Nanchang 330006, Jiangxi Province, China zhujunchi@tom.com
  • Supported by:

    the Youth Science Foundation of Department of Education of Jiangxi Province, No.GJJ09432*

PDF

501

被引次数

15

摘要：

背景：骨髓间充质干细胞是目前极具潜力的细胞和基因治疗的宿主细胞。体外标记是追踪其存活、“归巢”、生长、分化等一系列过程的前提。
目的：探索5-溴脱氧尿嘧啶核苷体外标记兔骨髓间充质干细胞的最佳时间和最佳浓度。
方法：兔骨髓间充质干细胞取自纯种健康新西兰大耳白兔，通过形态学及流式细胞术鉴定分离培养的细胞为骨髓间充质干细胞。观察不同5-溴脱氧尿嘧啶核苷浓度标记及不同5-溴脱氧尿嘧啶核苷标记时间对兔骨髓间充质干细胞的标记阳性率。
结果与结论：流式细胞术检测培养的细胞为较纯的骨髓间充质干细胞，生长曲线呈S形。5-溴脱氧尿嘧啶核苷标记的阳性细胞率随标记时间延长以及标记物浓度增加而逐渐增高, 40 μmol/L及72 h标记阳性率达85%~95%。结果提示，应用5-溴脱氧尿嘧啶核苷体外标记骨髓间充质干细胞是安全可靠的，体外标记最佳时间和最佳浓度分别为72 h和40 μmol/L。

关键词: 骨髓间充质干细胞, 5-溴脱氧尿嘧啶核苷, 流式细胞术, 兔, 全骨髓培养法

Abstract:

BACKGROUND: Currently, bone marrow mesenchymal stem cells (BMSCs) are potential cells and host cells for gene therapy. Labeling in vitro is a premise to track their survival, “homing”, growth, differentiation and so on.
OBJECTIVE: To explore the optimal 5-bromodeoxyuridine (BrdU) concentration and time of labeling for rabbit BMSCs in vitro.
METHODS: Rabbit BMSCs were harvested from bone marrow of purebred healthy New Zealand white rabbits. BMSCs were identified by flow cytometry and morphology. Rabbit BMSCs-labeling positive rates using BrdU at different concentrations and different incubation time were observed.
RESULTS AND CONCLUSION: Flow cytometry finally confirmed that the cells we cultured were BMSCs combined with morphological observation; the growth curve of rabbit MSCs was “S” shape. The positive rate of BMSCs labeled by BrdU increased with incubation time and BrdU concentration, and more than 85%-95% BMSCs were labeled after at optimal 40 μmol/L and 72 hours. Above-mentioned results have suggested that application of BrdU labeling for BMSCs in vitro is safe and reliable. The optimal time and concentration for labeling BMSCs are 72 hours and 40 μmol/L.

中图分类号: