摘要
目的:探讨建立喉部黏膜间充质干细胞的分离培养方法,并对其生物学特性进行鉴定,为进一步研究其在喉部瘢痕中形成的作用及喉部组织工程提供参考。方法:以比格犬喉部会厌背侧(舌面)黏膜为研究对象,采用消化培养的方法分离具有间充质干细胞样细胞。选取第3代细胞对其进行生物学特性鉴定,首先利用MTT法检测其增殖活性及克隆形成情况,然后通过流式细胞术检测细胞表面分子标记物CD29及CD34的表达情况,最后应用第3代细胞对其进行成脂肪细胞和成骨细胞分化培养,观察其分化能力。结果:分离培养细胞形态较为一致,绝大多数呈梭形,排列不规则。MTT增殖活性实验及克隆形成试验结果显示,所分离的细胞具有良好的增殖活性和克隆形成率;流式细胞术结果显示,该细胞表达CD29间充质干细胞细胞表面标记物,低表达造血干细胞细胞表面标记物CD34;同时,该细胞诱导分化成脂肪细胞和成骨细胞实验表明,其具有多向分化潜能。结论:从比格犬会厌背侧黏膜分离的细胞具有间充质干细胞样特性,为进一步研究喉部瘢痕形成机制及喉部组织工程提供了技术基础。
Objective： To establish the methods of isolation and cultivation of mesenchymal cells from laryngeal mucosa of beagle and investigate the biological characteristics of the isolated cells.Methods： Laryngeal mucosa were harvested by enzyme digestion.And the third passage of cells were collected to determine the clone formation ability and growth kinetics.And flow cytometry method was used to detect the expression of CD29 and CD34.Then the third passage of isolated cells was treated with adipogenic nutrient media and osteogenic nutrient media respectively.Results： After being subcultured,the sizes of isolated cells were similar,and the majority of them were spindle-shaped with disorderly arrangement.The flow cytometry results showed that the isolated cells expressed the marker CD29 of mesenchymal stem cells,but not CD34.After being cultured in inducing medias,the isolated cells could differentiate into adipocytes and osteoblasts.Conclusions： Mesenchymal stem cell-like cells existed in laryngeal mucosa of beagle,and these cells may be useful for the research of laryngeal keloid and laryngeal tissueen gineering.
出处
《现代生物医学进展》
CAS
2013年第15期2875-2878,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30973285)
关键词
喉部黏膜
分离培养
间充质干细胞
生物学特性
Laryngeal mucosa
Isolation and cultivation
Mesenchymal stem cell
Biological characteristics
