组织工程与重建外科杂志 ›› 2014, Vol. 10 ›› Issue (1): 11-13.doi: 10.3969/j.issn.1673-0364.2014.01.004

• 论著 • 上一篇    下一篇

软骨细胞生物打印后细胞活力分析

曲淼,沈聪聪,侯亦康,许佑荣,柴岗,高晓燕   

  1. 上海市上海交通大学医学院附属第九人民医院整复外科,上海市组织工程研究重点实验室;上海市上海市周浦医院
  • 发布日期:2020-07-23

Viability Assay on Biological Printing of Chondrocytes

QU Miao,SHEN Congcong,HOU Yikang,XU Yourong,CHAI Gang,GAO Xiaoyan   

  1. Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai Key Laboratory of Tissue Engineering;Shanghai Zhoupu Hospital
  • Published:2020-07-23
  • Contact: 国家自然科学基金项目(30600650);上海市自然基金资助项目(13zr1437500);2012年浦东新区卫生系统优秀青年医学人才培养基金资助项目(pwrq2012-14)

摘要: 目的初步确立软骨细胞的二维生物打印方法,实现对细胞喷射过程的控制并保持打印后的细胞活力。方法取原代软骨细胞,常规培养至第2代。实验分2组:打印组,快速成型组织打印机进行二维细胞打印,X轴间隔300μm,Y轴间隔1 500μm,激光共聚焦显微镜观察,经生物打印后培养2 h,Live/Dead viability Kit测定细胞活力,激光共聚焦显微镜观察细胞荧光染色情况;对照组除细胞悬液不行打印,其余操作同打印组。结果打印组细胞激光共聚焦显微镜观察,"细胞墨滴"在二维组织中均匀分布,满足二维设计细胞打印的要求,每个"细胞墨滴"含细胞15~35个。细胞活力测试显示,打印组细胞活力与对照组无明显区别。结论通过生物打印技术可实现软骨细胞在二维平面上的定向、定量规则分布,为进一步的细胞三维打印乃至器官打印体系奠定基础。

关键词: 软骨细胞, 细胞打印, 组织工程

Abstract: Objective To establish a two-dimensional biological printing technique of chondrocytes so as to control the cell transfer process and keep cell viability after printing. Methods Primary chondrocytes were obtained from auricles of 8-week-old piglets and then were regularly sub-cultured to passage 2 (P2). The experiment was divided into 2 groups:printing group and control group. In printing group, P2 chondrocytes were transferred by rapid prototype biological printer (interval in x-axis 300 μm, interval in y-axis 1 500 μm), and were then cultured for 2 hours, afterwards cell viability was detected by Live/Dead viability Kit and cell fluorescence was observed by laser scanning confocal microscope; In control group, all steps were the same as printing group except that cell suspension received no printing. Results Laser scanning confocal microscope observation on the cells in printing group revealed the “cell ink droplets”. They were distributed regularly and evenly in the two-dimensional layer and each contained 15-35 cells, meeting the requirement of designing two-dimensional cell printing. The cells in printing group went through cell viability test, laser scanning confocal microscope observation showed that it was no significant difference between the control group and the printing groups in terms of cell viability. Conclusion Biological printing technique can realize the oriented, quantificational and regular distribution of chondrocytes in the two-dimensional plane and lays the foundation for the construction of three-dimensional cell printing or even organ printing system.

Key words: Chondrocytes, Cell printing, Tissue engineering

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