中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (23): 4247-4250.doi: 10.3969/j.issn.1673-8225.2011.23.016

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

脂肪源性干细胞成骨分化过程中成骨相关miRNA的表达模式

张  浩1，康  焱1，马元琛2，张紫机1，黄保丁1，廖威明1   

  1. 1中山大学附属第一医院，广东省广州市   510000
    2广东省人民医院，广东省广州市    510000
  • 收稿日期:2011-03-29 修回日期:2011-05-06 出版日期:2011-06-04 发布日期:2011-06-04
  • 作者简介:张浩☆，男，1982年生，河南省安阳市人，汉族，中山大学附属第一医院关节外科在读博士，主要从事脂肪干细胞及骨组织工程研究。 zhanghao-sysu@163.com
  • 基金资助:

    国家自然科学基金(30872615)，名称：双层一体Collagen/TCP梯度软骨修复材料生物响应特性的机理研究；丹麦战略研究基金(2101-07-0120)，名称：Individualized Musculoskeletal Regeneration and Reconstruction Network。

Osteogenesis-specific miRNA expression pattern analysis in osteogenic differentiation of adipose-derived stem cells

Zhang Hao1, Kang Yan1, Ma Yuan-chen2, Zhang Zi-ji1, Huang Bao-ding1, Liao Wei-ming1   

  1. 1First Affiliated Hospital, Sun Yat-sen University, Guangzhou  510000, Guangdong Province, China
    2Guangdong Provincial People’s Hospital, Guangzhou  510000, Guangdong Province, China
  • Received:2011-03-29 Revised:2011-05-06 Online:2011-06-04 Published:2011-06-04
  • About author:Zhang Hao☆, Studying for doctorate, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China zhanghao-sysu@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30872615*; Denmark  Strategy Research Foundation, No. 2101-07-0120*

PDF

508

被引次数

4

摘要：

背景：miRNA在细胞分化等众多生理病理过程中起重要调控作用，可调节脂肪来源干细胞成骨分化。
目的：筛选成骨分化相关miRNA，分析miRNA在脂肪来源干细胞成骨分化过程中的表达模式。
方法：从皮下脂肪组织中分离、培养人脂肪来源干细胞。应用基因芯片技术筛选脂肪来源干细胞成骨诱导前后表达差异显著的miRNA，采用RT-PCR技术检测所筛选的miRNA在成骨诱导第7，14，21天上调/下调的相对表达强度，以ELISA试剂盒检测相应时间点成骨相关蛋白的表达情况。
结果与结论：①倒置显微镜下观察，传3代后可获得均一性较高的脂肪来源干细胞，一定诱导条件下具有成骨、成脂、成软骨分化能力。②基因芯片技术筛选出成骨分化前后表达差异变化明显的9个miRNA，其中5个上调，4个下调。③成骨诱导第7天，miR-106a表达上调1.58倍(P < 0.05)。第14天时，9个miRNA均表达上调。第21天时，5个miRNA表达上调，4个表达下调。④成骨相关蛋白骨钙素、碱性磷酸酶、Ⅰ型胶原及骨唾液酸蛋白的质量浓度在诱导成骨分化第7天即明显升高，第14天达到峰值，第21天略有下降。

关键词: 成骨分化, miRNA, 脂肪来源干细胞, 基因芯片, 表达模式

Abstract:

BACKGROUND: miRNAs have emerged as important regulators in various physiological and pathological processes of cell differentiation, and can regulate the osteogenic differentiation of adipose-derived stem cells.
OBJECTIVE: To screen the osteogenesis-specific miRNAs, and analyze the expression pattern of these miRNAs in osteogenic differentiation of adipose-derived stem cells. 
METHODS: Adipose-derived stem cells were isolated and cultured from human subcutaneous fat. The osteogenesis-specific miRNAs were screened by gene microarray technique. The relative expression of these miRNAs was analyzed on 7, 14, and 21 days by RT-PCR. The osteogenesis-specific proteins were detected on 7, 14, and 21 days by enzyme linked immunosorbent assay kit.
RESULTS AND CONCLUSION: ①The 3rd passage adipose-derived stem cells were homogeneous. Osteogenesis, adipogenesis, and chondrogenesis differentiation of adipose-derived stem cells need specific condition under an inverted microscope. ②Nine osteogenesis-specific miRNAs were picked up by gene microarray technique, five were upregulated and four were downregulated. ③On day 7 in osteogenic differentiation, miR-106a expression was upregulated 1.58 folds (P < 0.05). On day 14, nine miRNAs were upregulated. On day 21, five miRNAs were upregulated and four were downregulated. ④The concentration of osteogenesis-specific proteins such as osteocalcin, alkaline phosphatase, collagen Ⅰ and bone sialoprotein were increased on day 7, peaked on day 14 and slightly decreased on day 21. 

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