Cho Jung-Sun, Park Joo-Hoo, Kang Ju-Hyung, Kim Sung Eun, Park Il-Ho, Lee Heung-Man
Biomedical Science, College of Medicine, Korea University, Seoul 152-703, Korea College of Medicine, Institute for Medical Devices Clinical Trial Center, Guro Hospital, Korea University, Seoul 152-703, Korea.
Biomedical Science, College of Medicine, Korea University, Seoul 152-703, Korea.
Exp Biol Med (Maywood). 2015 Feb;240(2):185-93. doi: 10.1177/1535370214553898. Epub 2014 Oct 6.
Mesenchymal stem cells (MSCs) are multipotent progenitor cells in adult tissues. This study aimed to investigate nasal polyp (NP) tissues as a potential new source of multipotent MSCs that maintain their stemness and differentiation potential following multiple rounds of passaging. NP tissues were obtained from 10 patients during endoscopic sinus surgery. After isolating and culturing NP-derived MSCs (npMSCs), the expression levels of the surface markers CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD146 and human leukocyte antigens-class II DR antigen (HLA-DR) were estimated by flow cytometry. NpMSCs were cultured in chondrogenic, osteogenic, adipogenic, or neurogenic differentiation medium. The differentiation potential of npMSCs was analyzed by Alcian blue, alizarin red S, oil red O, and immunocytochemical staining and reverse transcription-polymerase chain reaction. The clonogenic potential of npMSCs was measured using a colony-forming unit assay. Cell proliferation of npMSCs was measured using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Flow cytometry analysis revealed that npMSCs were negative for hematopoietic lineage markers (CD34, CD45, and HLA-DR) and positive for MSC markers (CD44, CD73, CD90, and CD105). The npMSCs differentiated into osteogenic, adipogenic, chondrogenic, and neurogenic lineages, respectively. Chondrogenically differentiated npMSCs were stained with Alcian blue, osteogenically differentiated npMSCs were stained with alizarin red S, and adipogenically differentiated npMSCs were stained with oil red O. Real-time polymerase chain reaction results showed that the differentiated npMSCs expressed the respective differentiation markers (Sox 9 and Col2A for chondrogenesis, Runx2 and osteocalcin for osteogenesis, fatty acid-binding protein 4 and peroxisome proliferator-activated receptor γ for adipogenesis, TuJ1, neurofilament light chain, and neurofilament heavy chain for neurogenesis). There were no significant differences in the clonogenic potential and proliferation rate between early and late passage npMSCs. These results show that npMSCs possess the characteristics of MSCs in terms of morphology, multipotent differentiation capacity, cell surface marker expression, and clonogenicity. Thus, npMSCs may represent an alternative source of MSCs.
间充质干细胞（MSCs）是成体组织中的多能祖细胞。本研究旨在探究鼻息肉（NP）组织作为多能MSCs潜在新来源的可能性，这些MSCs在多轮传代后仍能保持其干性和分化潜能。在内镜鼻窦手术期间从10例患者获取NP组织。分离并培养NP来源的MSCs（npMSCs）后，通过流式细胞术评估表面标志物CD34、CD44、CD45、CD73、CD90、CD105、CD106、CD146和人类白细胞抗原II类DR抗原（HLA-DR）的表达水平。将npMSCs培养于软骨形成、成骨、脂肪形成或神经形成分化培养基中。通过阿尔辛蓝、茜素红S、油红O以及免疫细胞化学染色和逆转录-聚合酶链反应分析npMSCs的分化潜能。使用集落形成单位测定法测量npMSCs的克隆形成潜能。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法测量npMSCs的细胞增殖。流式细胞术分析显示，npMSCs对造血谱系标志物（CD34、CD45和HLA-DR）呈阴性，而对MSCs标志物（CD44、CD73、CD90和CD105）呈阳性。npMSCs分别分化为成骨、脂肪形成、软骨形成和神经形成谱系。软骨形成分化的npMSCs用阿尔辛蓝染色，成骨分化的npMSCs用茜素红S染色，脂肪形成分化的npMSCs用油红O染色。实时聚合酶链反应结果显示，分化的npMSCs表达各自的分化标志物（软骨形成的Sox 9和Col2A、成骨的Runx2和骨钙素、脂肪形成的脂肪酸结合蛋白4和过氧化物酶体增殖物激活受体γ、神经形成的TuJ1、神经丝轻链和神经丝重链）。早期和晚期传代的npMSCs在克隆形成潜能和增殖率方面无显著差异。这些结果表明，npMSCs在形态、多能分化能力、细胞表面标志物表达和克隆形成性方面具有MSCs的特征。因此，npMSCs可能代表MSCs的一种替代来源。
