中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (29): 4630-4635.doi: 10.3969/j.issn.2095-4344.2014.29.007

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

低氧对脂肪干细胞和关节软骨细胞三维共培养成软骨能力的影响

戴 兵，徐海艇，金海东，陈 辉，蔡建武，范时洋，潘 骏   

  1. 温州医科大学附属第二医院骨科，浙江省温州市 325027
  • 修回日期:2014-04-15 出版日期:2014-07-09 发布日期:2014-07-09
  • 通讯作者: 潘骏，教授，硕士生导师，温州医科大学附属第二医院骨科，浙江省温州市 325027
  • 作者简介:戴兵，1987年生，浙江省温州市人，汉族，温州医科大学在读硕士，主要从事创伤外科专业研究。
  • 基金资助:

    浙江省自然科学基金资助项目(Y2100899)；浙江省科技厅基金资助项目(2009C33131)；温州市科技局基金资助项目(H20080057)；温州市科技局基金资助项目(Y20100026)

Hypoxia effects on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes

Dai Bing, Xu Hai-ting, Jin Hai-dong, Chen Hui, Cai Jian-wu, Fan Shi-yang, Pan Jun   

  1. Department of Orthopedics, Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China
  • Revised:2014-04-15 Online:2014-07-09 Published:2014-07-09
  • Contact: Pan Jun, Professor, Master’s supervisor, Department of Orthopedics, Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China
  • About author:Dai Bing, Studying for master’s degree, Department of Orthopedics, Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China
  • Supported by:

    theNatural Science Foundation of Zhejiang Province, No. Y2100899; the Project of Science and Technology Department of Zhejiang Province, No. 2009C33131; the Project of Wenzhou Municipal Science and Technology Bureau, No. H20080057; the Project of Wenzhou Municipal Science and Technology Bureau, No. Y20100026

PDF

352

被引次数

8

摘要：

背景：多项体内外研究表明低氧和共培养均促进干细胞向软骨细胞方向分化。

目的：观察低氧对脂肪干细胞和关节软骨细胞三维共培养成软骨能力的影响。
方法：脂肪干细胞和关节软骨细胞二者按3∶1比例混合，以5×1010 L-1接种于聚乳酸-羟基乙酸共聚物/明胶支架上，分别在常氧(体积分数为20% O2)、低氧(体积分数为5% O2)环境下培养6周。苏木精-伊红染色进行组织学分析，阿尔新蓝染色鉴定糖胺多糖的合成，免疫组化鉴定Ⅱ型胶原的表达，并测定各组支架-细胞复合物的DNA、糖胺聚糖、羟脯氨酸含量。

结果与结论：低氧组苏木精-伊红染色显示大量细胞及细胞外基质生成，阿尔新蓝染色显示有大量糖胺多糖生成，免疫组化显示Ⅱ型胶原表达强阳性，且DNA、糖胺聚糖、羟脯氨酸等各项指标均高于常氧组。表明低氧促进脂肪干细胞和关节软骨细胞共培养成软骨分化。



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


全文链接：

关键词: 组织构建, 软骨细胞, 低氧, 脂肪干细胞, 聚乳酸-羟基乙酸共聚物, 共培养, 组织工程, 免疫组织化学, 浙江省自然科学基金

Abstract:

 BACKGROUND: Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes.

OBJECTIVE: To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes.
METHODS: Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×1010/L. The cells were cultured in normoxia (20% O2) and hypoxic (5% O2) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II collagen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cell complex was measured.

RESULTS AND CONCLUSION: In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix; alcian blue staining showed massive glycosaminoglycan formation; immunohistochemistry staining detected strongly positive expression of collagen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes.



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


全文链接：

Key words: stem cells, adipose tissue, chondrocytes, tissue scaffolds, co-culture techniques, cell hypoxia

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