1 Department of Otolaryngology-Head and Neck Surgery, Seoul St Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea.
2 Institute of Clinical Medicine Research, College of Medicine, The Catholic University of Korea, Seoul, Korea.
Otolaryngol Head Neck Surg. 2018 Oct;159(4):774-782. doi: 10.1177/0194599818777195. Epub 2018 May 22.
Objective To produce alternate cell sources for tissue regeneration, human nasal septal cartilage-derived progenitor cells (NSPs) were tested to identify whether these cells meet the criteria of cartilage progenitor cells. We also evaluated the effects of prolonged cultivation on the characteristics of NSPs. Study Design In vitro study. Setting Academic research laboratory. Methods NSPs were isolated from discarded human nasal septal cartilage. NSPs were cultured for 10 passages. The expression of septal progenitor cell surface markers was assessed by fluorescence-activated cell sorting. Cell proliferation was measured with a cell-counting kit. Cytokine secretion was analyzed with multiplex immunoassays. Chondrogenic differentiation of NSPs without differentiation induction was analyzed with type II collagen immunohistochemistry. Cartilage-specific protein expression was evaluated by Western blotting. Under osteo- and adipodifferentiation media, 2 lineage differentiation potentials were evaluated by histology and gene expression analysis. Results Surface epitope analysis revealed that NSPs are positive for mesenchymal stem cells markers and negative for hematopoietic cell markers. Cultured NSPs showed sufficient cell expansion and chondrogenic potential, as demonstrated by immunostaining and expression of cartilage-specific protein. IL-6, IL-8, and transforming growth factor ß were secreted by over 200 pg/mL. The osteo- and adipodifferentiation potentials of NSPs were identified by histology and specific gene expression. The aforementioned characteristics were not influenced by prolonged cultivation. Conclusion NSPs represent an initial step toward creating a cell source from surgically discarded tissue that may prove useful in cartilage regeneration.
目的 为了寻找组织再生的替代细胞来源,我们对人鼻中隔软骨衍生祖细胞(NSP)进行了测试,以确定这些细胞是否符合软骨祖细胞的标准。我们还评估了延长培养对 NSP 特性的影响。
研究设计 体外研究。
设置 学术研究实验室。
方法 从废弃的人鼻中隔软骨中分离 NSP。NSP 培养 10 代。通过流式细胞术评估鼻中隔祖细胞表面标志物的表达。用细胞计数试剂盒测量细胞增殖。通过多重免疫分析分析细胞因子分泌。通过 II 型胶原免疫组化分析未经诱导分化的 NSP 的软骨分化。通过 Western blot 评估软骨特异性蛋白的表达。在成骨和成脂分化培养基中,通过组织学和基因表达分析评估 2 种谱系分化潜能。
结果 表面表位分析表明,NSP 对间充质干细胞标志物呈阳性,对造血细胞标志物呈阴性。培养的 NSP 表现出足够的细胞扩增和软骨形成潜力,这通过免疫染色和软骨特异性蛋白的表达得到证实。IL-6、IL-8 和转化生长因子 β 的分泌量超过 200 pg/mL。NSP 的成骨和成脂分化潜能通过组织学和特定基因表达来鉴定。延长培养不会影响上述特征。
结论 NSP 是从手术切除的组织中创建细胞来源的初始步骤,可能对软骨再生有用。