中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7439-7442.doi: 10.3969/j.issn.1673-8225.2010.40.007

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

脐带间充质干细胞向成骨细胞分化的潜能

扎拉嘎胡1，陈  莉1，兰晓霞2，陈  晓3，陈小义1   

  1. 武警医学院，1细胞生物学教研室， 2预防医学系流行病学教研室，天津市300162；3武警医学院附属医院妇产科，天津市  300162
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 通讯作者: 陈小义，硕士，硕士生导师，教授，武警医学院细胞生物学教研室，天津市300162 chenxiaoyi6111@163.com
  • 作者简介:扎拉嘎胡★，女，1972年生，内蒙古自治区通辽市人，蒙古族，2003年内蒙古大学毕业，硕士，讲师，主要从事间充质干细胞基础和应用方面的研究。 zhalaga2003@sina.com
  • 基金资助:

    武警部队科研项目(WY200806)，课题名称：FAK介导信号通路与人脐血间充质干细胞成骨化作用。

Potential of human umbilical cord mesenchymal stem cells differentiating into osteoblasts

Zhala Ga-hu1, Chen Li1, Lan Xiao-xia2, Chen Xiao3, Chen Xiao-yi1   

  1. 1 Department of Cell Biology, Medical College of Chinese People’s Armed Police Forces, Tianjin  300162, China; 2 Research Room of Epidemiology, Department of Preclinical Medicine, Medical College of Chinese People’s Armed Police Forces, Tianjin  300162, China; 3 Department of Gynaecology and Obstetrics, Hospital Affiliated to Medical College of Chinese People’s Armed Police Forces, Tianjin  300162, China
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Chen Xiao-yi, Master, Master’s supervisor, Professor, Department of Cell Biology, Medical College of Chinese People’s Armed Police Forces, Tianjin 300162, China chenxiaoyi6111@163.com
  • About author:Zhala Ga-hu★, Master, Lecturer, Department of Cell Biology, Medical College of Chinese People’s Armed Police Forces, Tianjin 300162, China Zhalaga2003@sina.com
  • Supported by:

    the Scientific Research Program of Chinese People’s Armed Police Forces, No. WY200806*

PDF

355

被引次数

8

摘要：

背景：人脐带间充质干细胞含量丰富，较为原始，分化能力强，免疫原性低，是细胞治疗的理想靶细胞。
目的：体外分离培养脐带间充质干细胞并将其定向诱导为成骨细胞。
方法：无菌条件下培养脐带间充质干细胞，分为诱导组和对照组，诱导组用成骨诱导液处理、对照组为干细胞培养液处理。倒置光显微镜观察细胞形态，MTT法测细胞增殖，荧光双染法检测细胞活力，流式细胞仪检测细胞周期与细胞表面标记。诱导后：检测碱性磷酸酶，Von Kossa染色分析钙盐沉积，RT-PCR检测骨桥蛋白基因、碱性磷酸酶、骨唾液蛋白mRNA的表达。
结果与结论：传代细胞形态稳定、活力好，高标达CD44。诱导后von Kossa染色表现为阳性。碱性磷酸酶活性诱导组比对照组高(P < 0.05)，不同时间点比较21 d最高(P < 0.05)。RT-PCR显示：诱导组21，28d 碱性磷酸酶mRNA表达均较与对照组增强(P < 0.05)。诱导组有骨唾液蛋白和骨桥蛋白基因mRNA表达。提示，人脐带间充质干细胞能够定向分化为成骨细胞。

关键词: 脐带, 间充质干细胞, 成骨细胞, 细胞分化, 诱导

Abstract:

BACKGROUND: Human umbilical cord mesenchymal stem cells (UCMSCs) were rich and more primordial, had strong differentiation ability and low immunogenicity, suitable for transplanting to different individuals. Therefore, UCMSCs were an ideal target cells for cell therapy.
OBJECTIVE: To study the isolation and culture of UCMSCs and to explore the potential of osteogenesis differentiation in vitro. 
METHODS: UCMSCs were sterilely isolated and cultured. The third passage of cells were divided into induction group and control group. The induction group was treated with ossify induction medium, and control group was treated with stem cell culture medium. The cell morphology was observed under an inverted light microscope. Cell proliferation was determined by MTT assay. The cell viability was detected by double immunofluorescent staining. Cell cycle and cell surface marker were determined using flow cytometry. Following induction, alkaline phosphatase (ALP) activity was measured utilizing microplate reader. Calcium deposition was analyzed using Von Kossa staining. mRNA expression of osteopontin (OPN), ALP and bone sialoprotein (BSP) was measured by reverse transplantation-polymerase chain reaction (RT-PCR).
RESULTS AND CONCLUSION: The passage cells were steady in the cell shapes, with good viability, and highly expressed CD44. Following induction, the cells were positive in von Kossa staining. ALP activity was higher in induction group compared with control group (P < 0.05). The ALP activity was greatest on day 21 (P < 0.05). RT-PCR indicated that ALP mRNA expression was higher in induction group compared with control group (P < 0.05). The BSP and OPN genes were expressed in induction group, which suggested that human UCMSCs can differentiate into osteoblasts.

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