中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9131-.doi: 10.3969/j.issn.1673-8225.2010.49.003

• 干细胞培养与分化 • 上一篇    下一篇

骨髓间充质干细胞成骨方向诱导过程中的基因表达

胡杨，马莹，何惠宇，盛磊，阿布力孜•阿布杜拉，阿尔孜古丽   

  1. 新疆医科大学第一附属医院口腔修复科，新疆维吾尔自治区乌鲁木齐市  830054
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 何惠宇，博士，副教授，副主任医师，主任，新疆医科大学第一附属医院口腔修复科，新疆维吾尔自治区乌鲁木齐市 830054 joe98344@sina.cn
  • 基金资助:

    新疆维吾尔自治区科技攻关项目，项目名称：组织工程骨用于即刻牙种植骨结合的实验研究，项目编号：200533118；新疆维吾尔自治区高校科研计划科学研究重点项目，项目名称：bFGF基因转染的骨髓间充质干细胞复合异种煅烧骨修复颌骨缺损的实验研究，项目编号：xj EDU2009I22。

Gene expression during induced differentiation of bone marrow mesenchymal stem cells into osteoblasts

Hu Yang, Ma Ying, He Hui-yu, Sheng Lei, Abulizi•Abudula, Aerziguli   

  1. Department of Prosthodontics, First Affiliated Hospital, Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: He Hui-yu, Doctor, Associate professor, Associate chief physician, Department of Prosthodontics, First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China joe98344@sina.cn
  • Supported by:

    Key Scientific Program of Xinjiang Uygur Autonomous Region, No. 200533118*; the Scientific Research Key Program of Scientific Research Project of Higher Learning School of Xinjiang Uygur Autonomous Region, No.xjEDU2009I22*

PDF

536

被引次数

12

摘要：

背景：目前骨髓间充质干细胞向成骨细胞分化演变中的基因表达模式尚不明确。
目的：观察骨髓间充质干细胞向成骨细胞诱导分化过程中碱性磷酸酶、骨桥蛋白、Ⅰ型胶原、碱性成纤维细胞生长因子和骨钙素基因的表达情况，验证骨髓间充质干细胞是否向成骨细胞成熟分化。
方法：抽取2月龄新西兰大白兔股骨骨髓，全骨髓贴壁法培养获得骨髓间充质干细胞，用矿化诱导培养基(DMEM/F12、地塞米松1×10-8 mmol/L、β-磷酸甘油钠0.01 mol/L、维生素C 0.05 g/L)进行成骨诱导培养，用反转录-聚合酶链反应方法检测诱导培养后第一二代骨髓间充质干细胞碱性磷酸酶、骨桥蛋白、Ⅰ型胶原、碱性成纤维细胞生长因子和骨钙素基因的表达情况，并对该骨髓间充质干细胞表面抗原CD44进行鉴定。
结果与结论：经矿化诱导培养基诱导培养后，第一二代骨髓间充质干细胞阶段性表达碱性磷酸酶、骨桥蛋白、Ⅰ型胶原、碱性成纤维细胞生长因子和骨钙素基因；第1代骨髓间充质干细胞抗原CD44阳性率达44.4%。提示兔骨髓间充质干细胞在体外矿化诱导培养中逐渐向成骨细胞分化，分别于诱导后第一二代细胞中阶段性顺序表达成骨细胞特异性基因碱性磷酸酶、骨桥蛋白、Ⅰ型胶原、碱性成纤维细胞生长因子和骨钙素，该细胞已具备成骨细胞特征，为揭示骨髓间充质干细胞向成骨细胞分化过程中基因表达机制提供了实验依据。

关键词: 成骨细胞, 基因表达, 骨髓间充质干细胞, 分离培养, 诱导分化

Abstract:

BACKGROUND: At present, the gene expression mode of bone marrow mesenchymal stem cells (BMSCs) differentiation into osteoblasts remains unclear.
OBJECTIVE: To observe gene expression of alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin during BMSCs differentiation into osteoblasts, and to verify whether differentiation of BMSCs into osteoblasts is mature.
METHODS: The BMSCs were aspirated from the femoral bone of 2-month old New Zealand rabbits and cultured by all bone marrow adherence method, and induced to differentiate into osteoblasts in the medium supplemented with DMEM/F12, dexamethason 1×10-8 mmol/L, β-glycerophosphate sodium 0.01 mol/L and vitamin C 0.05 g/L. The gene expression of alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin in the first and second passages of BMSCs were examined by reverse transcription-polymerase chain reaction. BMSCs surface antigen CD44 was identified.
RESULTS AND CONCLUSION: Following induced culture in mineralized media, the first and second passages of BMSCs expressed alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin gene in some phases. Positive rate of CD44 in the first passage of BMSCs was 44.4%. These indicate that rabbit BMSCs gradually differentiated into osteoblasts following in vitro mineralization. The first and second passages of BMSCs expressed alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin in some phases. These cells have had the characteristics of osteoblasts and provided experimental evidences for revealing the mechanisms of gene expression of BMSCs differentiation into osteoblasts.

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