摘要
目的探讨不同年龄来源耳廓软骨的组织结构,软骨细胞的增殖、分化能力,以及相关基因和细胞表面标志物的表达差异。方法耳廓软骨组织来源为中国医学科学院整形外科医院收治的22例小耳畸形患者,年龄6~28岁,分为儿童组(6~12岁)、青少年组(13~18岁)和成人组(21~28岁)。提取得到不同年龄来源耳廓软骨细胞后,分别检测其增殖分化能力;同时应用荧光定量PCR检测细胞增殖和软骨细胞外基质相关基因在不同年龄软骨细胞中的表达差异;利用流式细胞术、免疫荧光、免疫组化等技术,检测间充质干细胞标志物CD90、CD44、CD73、CD105在这些细胞中的表达差异。结果儿童组耳廓软骨细胞的增殖能力较强,与成人组比较差异有统计学意义(P<0.05),与青少年组比较差异同样具有统计学意义(P<0.01);青少年和成人组软骨细胞增殖能力较弱;随着年龄的增加,软骨细胞外基质相关基因COL2A1的表达不断上升,儿童组与成人组比较P<0.01,青少年组与成人组比较P<0.01。细胞成骨分化能力随年龄的增长不断降低(P<0.05);但不同年龄来源软骨细胞的成脂分化能力比较差异无统计学意义;流式和荧光定量PCR的结果均表明间充质干细胞标志物的表达随着年龄的增长逐渐降低,以CD90最为明显(P<0.01)。结论年龄因素影响耳廓软骨组织来源细胞的生物学特性及干细胞含量。
Objective To investigate the tissue structure, chondrocyte characteristics, and the differential expression of related genes and cell surface markers of auricular cartilage of patients in different ages, in order to provide a basis for the age selection of tissue engineered cartilage repair defects. Methods The auricular cartilage tissue was obtained from 22 patients with microtia in the Plastic Surgery Hospital, Chinese Academy of Medical Science, ranged from 6 to 28 years old, and divided into the child group (6-12 years old), the adolescent group (13-18 years old) and the adult group (21-28 years old). The proliferation and differentiation features of chondrocytes which from different-aged patients were detected. Furthermore, quantitative real-time PCR was used to detect the differences in the expression of genes related to cell proliferation and chondrocyte extracellular matrix. Flow cytometry, immunofluorescence and immunohistochemistry were used to detect the differences in the expression of mesenchymal stem cell markers CD90, CD44, CD73 and CD105 in chondrocytes. SPSS Statistics 21.0 software was used to process statistics. Results The proliferation capability of auricular chondrocytes of children was stronger than adolescents and adults, the child group vs the adult group P<0.05, the child group vs the adolescent group P<0.01. The expression of cartilage extracellular matrix related gene COL2A1 increased with age, the child group vs the adult group P<0.01, the adolescent group vs the adult group P<0.01. While the capability of cell osteogenic differentiation decreased with age(P<0.05). However, there was no significant difference in the capability of adipogenic differentiation when considering the ages of patients. The results of both flow cytometry and real-time PCR showed that the expression of mesenchymal stem cell markers decreased with age, with the most significant decrease in CD90(P<0.01). Conclusions The biological characteristics and stem cell content of cells derived from auricular cartilage tis
作者
吴怡
董可欣
杨峥
刘霞
肖苒
蒋海越
Wu Yi;Dong Kexin;Yang Zheng;Liu Xia;Xiao Ran;Jiang Haiyue(Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100144, China)
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2019年第4期331-340,共10页
Chinese Journal of Plastic Surgery
基金
国家自然科学基金(81471804,81871575)
中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-007).
关键词
年龄
耳廓软骨
细胞增殖
细胞分化
软骨干细胞/祖细胞
Age
Auricular cartilage
Cell proliferation
Cell differentiation
Cartilage stem/progenitor cells
