中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (51): 7628-7633.doi: 10.3969/j.issn.2095-4344.2016.51.004

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

生长分化因子5诱导脂肪干细胞向软骨细胞的转化

李明辉,刘  洋,孙  凯,谭俊峰,张  觅   

  1. 江汉大学附属第二医院,武汉市第五医院骨科,湖北省武汉市  430050
  • 收稿日期:2016-10-06 出版日期:2016-12-09 发布日期:2016-12-09
  • 通讯作者: 刘洋,主任医师,三级教授,江汉大学附属第二医院,武汉市第五医院骨科,湖北省武汉市 430050
  • 作者简介:李明辉,1976年生,湖南省邵阳市人,汉族,2013年武汉大学医学院毕业,博士,副主任医师,主要从事运动创伤、关节疾患、四肢修复重建等方面的研究。
  • 基金资助:

    武汉市科技局科学技术计划资助项目(2013060602010265)

Growth differentiation factor 5 induces adipose-derived stem cells differentiating into chondrocytes

Li Ming-hui, Liu Yang, Sun Kai, Tan Jun-feng, Zhang Mi   

  1. Department of Orthopaedics, Fifth Hospital of Wuhan & the Second Affiliated Hospital of Jianghan University, Wuhan 430050, Hubei Province, China
  • Received:2016-10-06 Online:2016-12-09 Published:2016-12-09
  • Contact: Liu Yang, Chief physician, Professor, Department of Orthopaedics, Fifth Hospital of Wuhan & the Second Affiliated Hospital of Jianghan University, Wuhan 430050, Hubei Province, China
  • About author:Li Ming-hui, M.D., Associate chief physician, Department of Orthopaedics, Fifth Hospital of Wuhan & the Second Affiliated Hospital of Jianghan University, Wuhan 430050, Hubei Province, China
  • Supported by:

    the Science and Technology Program of the Wuhan Science and Technology Bureau, No. 2013060602010265

摘要:

文章快速阅读:

文题释义:
生长分化因子5:属骨形态发生蛋白家族成员,也是转化生长因子β超家族一员,属于转化生长因子β超家族的新亚族。生长分化因子5 已被多数学者证实具有促进成骨、软骨组织分化,促进肌腱、韧带、椎间盘及中枢、外周神经修复与发育,甚至对于心血管系统都发挥着重要的生理作用。在胚胎软骨生成的早期,生长分化因子5在关节表面和关节间隙的表达特别明显,介导软骨前体细胞的分化。
生长分化因子5的特性:可诱导多能干细胞向软骨、肌腱、骨等组织分化,离体条件下可诱导骨髓间充质干细胞分化为软骨。有研究发现,在间充质干细胞培养液中加入生长分化因子5 时,可显著促进间充质干细胞的增殖,并刺激软骨细胞和成骨细胞标志物的表达。生长分化因子5在诱导软骨分化上与转化生长因子β有协同作用,生长分化因子5 还参与骨与软骨损伤的修复。在关节软骨损伤时,可见大量生长分化因子5表达,说明生长分化因子5也参与软骨损伤的修复过程。
摘要
背景
:研究表明,生长分化因子5通过不同的调控机制在软骨发育中起到非常重要的作用。
目的:观察生长分化因子5对脂肪干细胞分化的调控作用。
方法:取第4代日本大耳白兔脂肪干细胞,分别以含0(空白对照组),10,50,100,150,200 μg/L生长分化因子5的培养液培养,观察细胞形态变化,筛选出最佳生长分化因子5质量浓度。取最佳质量浓度生长分化因子5诱导培养1,2,3周的细胞爬片,分别进行免疫组织化学、甲苯胺蓝及免疫荧光染色。
结果与结论:①最佳质量浓度:当生长分化因子5质量浓度为10,50 μg/L时,细胞形态变化不明显;在200 μg/L时,细胞呈现凋亡表现;在100,150 μg/L质量浓度下,可见少量细胞由梭形变得不规则,部分细胞在7-10 d变成圆形或椭圆形,其中以100 μg/L 生长分化因子5诱导效果最好;②免疫组织化学染色:100 μg/L生长分化因子5诱导后,转染细胞的Ⅱ型胶原化学染色显示细胞核蓝染,呈强阳性;③甲苯胺蓝染色:100 μg/L生长分化因子5诱导后,转染细胞的细胞甲苯胺蓝染色阳性,胞浆异染;④免疫荧光染色:100 μg/L生长分化因子5诱导组的蛋白聚糖绿色荧光表达逐渐增强,以诱导3周最为强烈;⑤结果表明:长分化因子5可促进脂肪干细胞向软骨细胞转化。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-3689-7011(刘洋)

关键词: 组织构建, 软骨细胞, 生长分化因子5, 脂肪干细胞, 诱导, 分化

Abstract:

BACKGROUND: Growth differentiation factor 5 (GDF5) has been shown to play a crucial role in the development of chondrocytes via different regulatory mechanisms.
OBJECTIVE: To explore the effect of GDF5 on the chondrogenic differentiation of adipose-derived stem cells (ADSCs).
METHODS: ADSCs in passage 4 isolated from Japanese White rabbits were cultured in the medium containing 0 (blank control group), 10, 50, 100, 150 and 200 μg/L, respectively. The morphological changes were observed, and the optimal concentration of GDF-5 was screened, and its round coverslips at 1, 2 and 3 weeks of culture were selected to undergo immunocytochemistry, toluidine blue staining and immunofluorescent staining.
RESULTS AND CONCLUSION: The growth of ADSCs was stable in the medium containing 10 and 50 μg/L GDF5, while apoptotic cells appeared after induction with 200 μg/L GDF5. In the medium containing 100 and 150 μg/L GDF5, some spindle-shaped cells changed into irregular shape, and became round or oval after 7-10 days of culture. The optimal concentration of GDF5 was 100 μg/L. The cells transfected by 100 μg/L GDF5 were positive for type II collagen obviously and with blue-stained nucleus. The transfected cells were positive for toluidine blue, and metachromatic granules were visible in the cytoplasm. The proteoglycan mRNA expression of the transfected cells was significantly increased, and reached the highest at 3 weeks. These results suggest that GDF5 promotes the chondrocytic differentiation of ADSCs.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Growth Differentiation Factors, Stem Cells, Chondrocytes, Tissue Engineering

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