组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2019, Vol. 15 ›› Issue (2): 81-84.doi: 10.3969/j.issn.1673-0364.2019.02.005

• 论著 • 上一篇    下一篇

左旋聚乳酸多孔微球体外动态培养耳郭软骨细胞的研究

孙恒，蒋海越，马世泽，刘霞，滕利   

  1. 中国医学科学院北京协和医学院整形外科医院
  • 收稿日期:2019-03-01 修回日期:2019-03-26 出版日期:2019-04-20 发布日期:2019-04-20
  • 作者简介:刘霞(E-mail:lacey2003@163.com);滕利(E-mail:tenglidr@sina.com);

Dynamic Culture of Auricular Chondrocytes with L-Polylactic Acid Porous Microspheres in Vitro

SUN Heng,JIANG Haiyue,MA Shize,LIU Xia,TENG Li   

  1. Plastic Surgery Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100144,China)
  • Received:2019-03-01 Revised:2019-03-26 Online:2019-04-20 Published:2019-04-20
  • Contact: 国家自然科学基金(81471804)；中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-007)

摘要： 目的 明确PLLA(左旋聚乳酸)、Cultispher G两种多孔微球作为耳郭软骨细胞体外培养载体的可行性,为软骨细胞三维动态培养寻找合适的微球支架。方法 分离猪耳郭软骨、培养软骨细胞,将获得的第1代软骨细胞分别接种于PLLA和Cultispher G多孔微球上,并于RCCS三维培养系统中进行培养。应用电镜和荧光倒置显微镜观察细胞在微球上的生长情况,CCK-8法检测软骨细胞增殖情况,Real-time PCR检测相关基因表达。结果 软骨细胞可贴附于两种微球表面,PLLA和Cultispher G两种微球接种率分别为(37.67%±2.33%)和(73.67%±3.53%)。培养21 d后,PLLA多孔微球细胞总数高于Cultispher G多孔微球。Real-time PCR检测发现PLLA多孔微球维持软骨细胞表型能力优于Cutispher G多孔微球。结论 与Cultispher G多孔微球相比,PLLA多孔微球更适合软骨细胞体外三维培养。

关键词: 左旋聚乳酸, 多孔微球, 三维培养

Abstract: Objective To clarify the feasibility of PLLA and Cultispher G porous microspheres as an in vitro culture material for auricular chondrocytes,and to find a suitable microsphere scaffold for the three-dimensional dynamic culture of chondrocytes.Methods Aricular chondrocytes were isolated and purified from swine ear.P1 chondrocytes were inoculated on PLLA microspheres and Cultispher G microspheres,then cultured in a rotary cell culture system(RCCS).Growth of chondrocytes was observed on the microcarriers by using inverted and scanning electron microscopes.Proliferation rate of chondrocyte was measured by CCK-8 examination method.Relevant genes were measured by real-time PCR.Results Inverted and scanning electron microscopes showed chondrocytes could grow on PLLA microspheres and Cultispher G microspheres.Inoculation rates of chondrocytes were(37.67%±2.33%)and(73.67%±3.53%)respectively.Increasing fold of chondrocytes on PLLA microspheres was higher than that on Cultispher G microspheres after cultured for 21 days.Real-time PCR showed that PLLA microspheres maintained chondrocyte phenotype better than Cultispher G microspheres.Conclusion Compared with Cultispher G microsphere,PLLA porous microspheres are more suitable for three-dimensional culture of chondrocytes in vitro.

Key words: L-polylactide, Porous microsphere, Three-dimensional cultivation

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