학술논문
基于人诱导多能干细胞的组织工程化软骨再生 / Tissue-engineered cartilage regeneration based on human induced pluripotent stem cells
Document Type
Academic Journal
Author
Source
口腔颌面外科杂志 / Journal of Oral and Maxillofacial Surgery. 34(1):22-28
Subject
Language
Chinese
ISSN
1005-4979
Abstract
目的:构建一种稳定高效的人诱导多能干细胞(induced pluripotent stem cells,iPSCs)向软骨中胚层方向分化的培养方法,初步探究hiPSCs来源的软骨用于关节软骨再生的可能.方法:通过模拟体内发育过程和三维(three dimensions,3D)悬浮培养体系,逐步诱导hiPSCs向软骨中胚层和成熟的软骨组织分化;利用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)、蛋白质印迹法(Western blotting)、免疫荧光及免疫组织化学技术检测整个培养过程中多能干性、中胚层及软骨相关基因和蛋白的动态变化;最后利用裸鼠皮下模型研究hiPSCs软骨球软骨表型的稳定性及成瘤性.结果:通过模拟体内发育过程,成功逐步诱导hiPSCs向软骨中胚层方向分化,最终分化为成熟的软骨组织.结论:本研究成功构建了一种稳定高效诱导hiPSCs向软骨中胚层方向分化的培养体系,为探究hiPSCs介导的软骨组织发育和再生提供了一个研究平台.
Objective:To establish a stable and efficient culture approach for guiding human induced pluripotent stem cells(hiPSCs)differentiation towards chondrogenic mesodermal lineage,and preliminarily explore the possibility of hiPSCs-derived cartilaginous pellets for articular cartilage regeneration.Methods:By simulating the development process in vivo,hiPSCs were gradually induced to differentiate into chondrogenic mesodermal lineage and mature cartilage using three dimensions(3D)suspension culture system.Real-time quantitative polymerase chain reaction(RT-qPCR),Western blotting,immunofluorescence and immunohistochemistry were used to investigate the dynamic expression change of pluripotent,mesodermal and chondrongenic related genes and proteins during the whole culture procedure.Finally,subcutaneous model of nude mice was used to investigate the stability of chondrogenic phenotypes and tumorigenicity of hiPSCs derived-cartilaginous pellets.Results:By simulating the development process in vivo,hiPSCs were gradually induced to differentiate into chondrogenic mesodermal lineage,and finally differentiated into mature cartilage tissues.Conclusion:In this study,a stable and efficient culture approach for inducing hiPSCs differentiation towards chondrogenic mesoderm was successfully established,which provide a research platform for investigation of hiPSCs-mediated cartilage development and regeneration.
Objective:To establish a stable and efficient culture approach for guiding human induced pluripotent stem cells(hiPSCs)differentiation towards chondrogenic mesodermal lineage,and preliminarily explore the possibility of hiPSCs-derived cartilaginous pellets for articular cartilage regeneration.Methods:By simulating the development process in vivo,hiPSCs were gradually induced to differentiate into chondrogenic mesodermal lineage and mature cartilage using three dimensions(3D)suspension culture system.Real-time quantitative polymerase chain reaction(RT-qPCR),Western blotting,immunofluorescence and immunohistochemistry were used to investigate the dynamic expression change of pluripotent,mesodermal and chondrongenic related genes and proteins during the whole culture procedure.Finally,subcutaneous model of nude mice was used to investigate the stability of chondrogenic phenotypes and tumorigenicity of hiPSCs derived-cartilaginous pellets.Results:By simulating the development process in vivo,hiPSCs were gradually induced to differentiate into chondrogenic mesodermal lineage,and finally differentiated into mature cartilage tissues.Conclusion:In this study,a stable and efficient culture approach for inducing hiPSCs differentiation towards chondrogenic mesoderm was successfully established,which provide a research platform for investigation of hiPSCs-mediated cartilage development and regeneration.