中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (10): 1731-1735.doi: 10.3969/j.issn.1673-8225.2011.10.005

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

年龄对人骨髓间充质干细胞体外增殖能力的影响

王小艳1，王志伟2，王青山1，谢东方1，陈伯华3   

  1. 青岛市中心医院，1急诊科，2普外科，山东省青岛市 266042；3青岛大学医学院附属医院脊柱外科，山东省青岛市  266000
  • 收稿日期:2010-10-19 修回日期:2011-02-13 出版日期:2011-03-05 发布日期:2011-03-05
  • 通讯作者: 陈伯华，主任医师，青岛大学医学院附属医院脊柱外科，山东省青岛市 266000 bhchen@hotmail.com
  • 作者简介:王小艳☆，女，1980年生，山东省威海市人，汉族，2009年德国海德堡大学毕业，博士，医师，主要从事骨髓间充质干细胞的研究。 xiaoyan.wang445@hotmail.com

Effect of aging on proliferative capacity of human bone marrow mesenchymal stem cells in vitro

Wang Xiao-yan1, Wang Zhi-wei2, Wang Qing-shan1, Xie Dong-fang1, Chen Bo-hua3   

  1. 1Department of Emergency, 2Department of General Surgery, Qingdao Central Hospital, Qingdao  266042, Shandong Province, China; 3Department of Spinal Surgery, Affiliated Hospital of Qingdao University Medical College, Qingdao  266042, Shandong Province, China
  • Received:2010-10-19 Revised:2011-02-13 Online:2011-03-05 Published:2011-03-05
  • Contact: Chen Bo-hua, Chief physician, Department of Spinal Surgery, Affiliated Hospital of Qingdao University Medical College, Qingdao 266042, Shandong Province, China bhchen@hotmail.com
  • About author:Wang Xiao-yan☆, Doctor, Physician, Department of Emergency, Qingdao Central Hospital, Qingdao 266042, Shandong Province, China xiaoyan.wang445@hotmail.com

PDF

313

被引次数

10

摘要：

背景：人骨髓间充质干细胞的增殖和分化可以修复组织器官损伤，随着年龄增长人体组织修复能力减弱，是否人骨髓间充质干细胞增殖能力随年龄增长而减弱尚不清楚。
目的：观察年龄对人骨髓间充质干细胞体外增殖能力的影响。
方法：15份骨髓标本来源于青岛市中心医院骨折或行髋关节置换患者，其中儿童、成年人、老年人各5例，年龄分别为7~12岁，20~55岁，60~90岁。抽取人骨髓组织，采用密度梯度离心法分离出单核细胞并接种于培养瓶内，在第5天计数集落形成单位；当贴壁细胞融合后进行传代，第1代细胞分两种方式进行培养：192个细胞接种于2个96孔板观察单细胞克隆形成能力，12 500个细胞以5×103/cm2接种于培养瓶内并以此密度连续传代观察细胞增殖率。
结果与结论：不同年龄人群骨髓间充质干细胞体外培养集落形成单位和细胞增殖率无明显区别(P > 0.05)，但单细胞克隆形成能力随年龄增长而下降(P  < 0.01)。提示骨髓间充质干细胞的数量和增殖能力没有随年龄的增长而下降，但其中能形成单细胞克隆未定向干细胞的数量随年龄的增长而减少。

关键词: 年龄, 骨髓间充质干细胞, 体外, 增殖能力, 单克隆形成

Abstract:

BACKGROUND: Mesenchymal stem cells (MSCs) from human beings can proliferate and differentiate to repair the damages of tissues and organs. The reparation capacity of tissues and organs decreases with age. Question arises if the proliferation capacity of MSCs deceases with age.
OBJECTIVE: To evaluate the effect of aging on the proliferative capacity of human MSCs in vitro.
METHODS: Fifteen bone marrow specimens were obtained from the patients in Qingdao Central Hospital, who received the operation of bone because of bone fracture or hip diseases. They were divided into three groups: 5 children aged from 7 to 12 years; 5 adults aged from 20 to 55 years; 5 elder aged from 60 to 90 years. The bone marrow isolates were taken out from patients during operation. The mononuclear cells were isolated from bone marrow using density gradient centrifugation and seeded into the culture flask. On the fifth day, colony forming units were counted under microscope. When the cells reached a confluence, they were passaged and further culture was preceded in two ways: 192 cells were seeded in two 96-well plates to study the cloning efficiency of single cells; 12 500 cells were seeded in T25 flask at the density of 5×103/cm to study the proliferation rate of cells.
RESULTS AND CONCLUSION: There were no correlation between colony forming units and proliferation rate of MSCs cultured in vitro and the age (P > 0.05), but the cloning efficiency of single cells decreased with age (P < 0.01). The number and proliferation capacity of MSCs do not decrease with age, but the uncommitted stem cells that could proliferate to single cell clones do deplete with age.

中图分类号: