中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (19): 3563-3566.doi: 10.3969/j.issn.1673-8225.2011.19.034

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

鹿茸多肽诱导骨髓间充质干细胞体外向软骨表型的分化

修忠标1，林建华2，吴朝阳2，王日雄2   

  1. 1福建中医药大学附属人民医院骨伤科，福建省福州市   350004
    2福建医科大学附属第一医院骨科，福建省福州市  350005
  • 收稿日期:2010-11-04 修回日期:2010-12-19 出版日期:2011-05-07 发布日期:2011-05-07
  • 作者简介:修忠标☆，男，1966年生，辽宁省营口市人，汉族，2005年福建中医药大学毕业，博士，主任医师，硕士生导师，主要从事骨性关节炎的基础和临床研究。
  • 基金资助:

    福建省自然科学基金计划高校专项项目(C0740003)。

Effect of pilose antler polypeptides on chondrogenic phenotype differentiation of bone marrow-derived mesenchymal stem cells in vitro

Xiu Zhong-biao1, Lin Jian-hua2, Wu Zhao-yang2, Wang Ri-xiong2   

  1. 1Department of Orthopedics and Traumatology, the Affiliated People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou  350004, Fujian Province, China
    2Department of Orthopedics, the First Affiliated Hospital of Fujian Medical University, Fuzhou  350005, Fujian Province, China
  • Received:2010-11-04 Revised:2010-12-19 Online:2011-05-07 Published:2011-05-07
  • About author:Xiu Zhong-biao☆, Doctor, Chief physician, Master’s supervisor, Department of Orthopedics and Traumatology, the Affiliated People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou 350004, Fujian Province, China xzbdoctor@sina.com
  • Supported by:

    the Natural Science Foundation of Fujian Province, No. C0740003*

PDF

325

被引次数

7

摘要：

背景：实验证实鹿茸多肽可以促进体外培养软骨细胞的增殖和细胞外基质糖胺多糖、Ⅱ型胶原、Aggrecan蛋白的表达。
目的：通过对体外培养的兔骨髓间充质干细胞在特定培养液作用下向软骨细胞表型分化的研究，探讨鹿茸多肽对其软骨分化的影响。
方法：将第3代兔骨髓间充质干细胞随机分为空白对照组、诱导组、鹿茸多肽组，分别采用普通培养液、诱导培养液、含10 mg/L鹿茸多肽的诱导培养液于离心管内进行培养；并取兔的关节软骨细胞作为关节软骨组。分别于1，2，3周后取材，通过组织学、生物化学和RT-PCR技术，对离心管内构建的软骨组织进行形态学和细胞功能状态的观察。
结果与结论：空白对照组培养2周后，细胞团块逐渐崩解，无法进行苏木精-伊红染色。诱导组、鹿茸多肽组细胞团块除有轻度收缩外，呈白色半透明状；苏木精-伊红染色发现部分细胞为圆形或卵圆形，表层细胞密度大；诱导组、鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达随培养时间延长而增多，各时间点诱导组、鹿茸多肽组含量均高于空白对照组(P < 0.05)；各时间点鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达均高于诱导组，但低于关节软骨组 (P< 0.05)。提示骨髓间充质干细胞在特定培养条件下能向软骨细胞表型分化，且鹿茸多肽对其定向软骨分化有明显促进作用。虽然在体外可以构建出软骨组织，但其与关节软骨质量相比仍有很大差距。

关键词: 鹿茸多肽, 间质干细胞, 软骨, 组织工程, 分化

Abstract:

BACKGROUND: Pilose antler polypeptides (PAP) have been proved to promote the proliferation of condrocytes cultured in vitro and expressions of glycosaminoglycan (GAG), type Ⅱ collagen and Aggrecan protein in the extracellular matrix.
OBJECTIVE: To investigate the feasibility of chondrogenic phenotype differentiation of rabbit bone marrow-derived mensenchymal stem cells (BMSCs) in a defined medium and then to study the effect of PAP on chondrogenic differentiation of BMSCs in vitro.
METHODS: The third passage BMSCs from rabbits were randomly divided into control group cultured in ordinary medium, induced group cultured in defined medium, and PAP group cultured in defined medium containing 10 mg/L PAP. An equal volume of articular chondrocytes were selected from rabbits as articular cartilage group. The cellular morphological and functional characteristics were observed after 1, 2, 3 weeks in centrifuge tubes by histological, biochemical and reverse transcription-polymerase chain reaction (RT-PCR) technique.
RESULTS AND CONCLUSION: Cell masses in the control group gradually crumbled after 2 weeks, and hematoxylin-eosin staining could not be done. Cell masses in the induced and PAP groups were semitransparent, but slightly contracted. A part of these cells were round or oval with a high density distribution at the surface. The content of GAG and mRNA expression of type Ⅱ collagen in the induced and PAP groups were increased with culture time, and higher than those in the control group at different time points (P < 0.05). The content of GAG and mRNA expression of type Ⅱ collagen in the PAP group were higher than those in the induced group, but lower than those in the articular cartilage group (P < 0.05). The results indicated that BMSCs can differentiate into chondrogenic phenotype in the defined medium, and PAP can significantly enhance chondrogenic phenotype differentiation of BMSCs. But the quality of cultured cartilage tissue is poorer than that of the articular cartilage.

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