中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5891-5896.doi: 10.3969/j.issn.1673-8225.2010.32.001

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

富血小板血浆诱导兔骨髓基质干细胞向成骨细胞的分化

张黎龙1，路  磊2，张学利1 ,田  融1   

  1. 1天津市人民医院脊柱外科，天津市 300121；2中国医科大学绍兴华宇医院骨科，浙江省绍兴市 312030
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 作者简介:张黎龙★，男，1976年生，辽宁省凤城市人，汉族，1999年中国医科大学毕业，硕士，主治医师，主要从事脊柱外科方面的研究。zhang-lilong@medmail.com.cn

Platelet-rich plasma induces the differentiation of rabbit bone marrow stromal stem cells into osteoblasts

Zhang Li-long1, Lu Lei2, Zhang Xue-li1, Tian Rong1   

  1. 1 Department of Spinal Surgery, Tianjin People’s Hospital, Tianjin  300121, China; 2 Department of Orthopaedics, Shaoxing Huayu Hospital, China Medical University, Shaoxing  312030, Zhejiang Province, China
  • Online:2010-08-06 Published:2010-08-06
  • About author:Zhang Li-long★,Master, Attending physician,Department of Spinal Surgery, Tianjin People’s Hospital, Tianjin 300121, China zhang-lilong@medmail.com.cn

PDF

392

被引次数

20

摘要：

背景：对于骨髓基质干细胞向成骨细胞的定向诱导分化，目前大多存在诱导物价格昂贵、制备困难或细胞培养周期长、成骨能力低等缺点，近年研究发现浓缩血小板中存在的生长因子有诱导骨再生的作用。
目的：观察富血小板血浆对体外培养兔骨髓基质干细胞的增殖和成骨细胞分化作用。
方法：从8只兔双侧股骨大转子抽取骨髓，体外分离培养兔骨髓基质干细胞，传至第3代分为两组，实验组用含1%富血小板血浆的DMEM进行干预，对照组加入普通DMEM条件培养液。
结果与结论：倒置显微镜下，原代培养细胞24~36 h后开始贴壁，10~12 d细胞融合成单层。实验组细胞培养第2天已贴壁，第6天细胞大部分融合；对照组细胞形态学变化较实验组晚出现1~3 d。培养第2，6，10，14天，碱性磷酸酶活性测定和茜素红钙结节染色结果显示，体外培养的兔成骨细胞生长良好，生化指标稳定，表现出与典型的成骨细胞相似的形态特征和生物学特性，证实富血小板血浆能促进体外培养的兔骨髓基质干细胞增殖，并能促使其向成骨细胞分化。

关键词: 细胞分化, 富血小板血浆, 碱性磷酸酶, 成骨细胞, 骨髓基质干细胞

Abstract:

BACKGROUND: The directional induced differentiation of bone marrow stromal stem cells (BMSCs) into osteoblasts has disadvantages of high inducer price, difficult to prepare or long cell culture cycle and low osteogenic ability. Previous studies have confirmed that growth factor in concentrated platelet can induce osteanagenesis.
OBJECTIVE: To observe effects of platelet-rich plasma (PRP) on the proliferation and osteoblast differentiation activity of rabbit BMSCs cultured in vitro.
METHODS: Bone marrow was taken from the greater trochanter of bilateral femur in 8 rabbits. The rabbit BMSCs were separated and cultivated in vitro. The third generation of BMSCs was divided into two groups. In the experimental group, BMSCs were interfused with DMEM containing 1% PRP, while in the control group, BMSCs were incubated in common DMEM conditioned medium.
RESULTS AND CONCLUSION: Under an inverted microscope, the primary cells began to adhere to the wall at 24-36 hours. The cells were confluent at 10 -12 days. Cells in experimental group began to adhere on day 2. Most of the cells were confluent on day 6. The shape change of cells from the control group was later 1-3 days than that of the experimental group. At 2, 6, 10 and 14 days, alkaline phosphatase determination and the calcium node alizarin red staining showed that the rabbit osteobalasts cultured in vitro grew well by this method and the biochemical indexes were stable. They also had the same morphological and biological characters as the osteoblasts. These verified that the PRP can promote the BMSCs proliferation, and can also accelerate effectively the differentiation from BMSCs into osteoblasts.

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