Ishimura Daisuke, Yamamoto Naoki, Tajima Kaori, Ohno Ayumi, Yamamoto Yasuhiro, Washimi Osuke, Yamada Harumoto
Department of Orthopaedic Surgery, School of Medicine, Fujita Health University, Aichi, Japan.
Tohoku J Exp Med. 2008 Oct;216(2):149-56. doi: 10.1620/tjem.216.149.
The incidence of arthritic diseases is rapidly increasing in most advanced countries. Articular cartilage, which is the most important tissue in the joint, consists of chondrocytes and abundant extracellular matrix, including aggrecan, and shows poor self-repair. We studied the potential of stem cells in mouse subcutaneous adipose tissue as a source of cells to regenerate cartilage tissue. Analysis of adipose-derived stromal vascular fraction culture cells (ADSVFs) using mesenchymal stem cell markers showed that CD90-positive cells accounted for 93.8%, CD105-positive cells for 68.5%, and p75 neurotrophin receptor (p75NTR, CD271)-positive cells for 36.1%. These results indicate that cells positive for mesenchymal stem cell markers are present in ADSVFs. The CD105-positive or -negative cells were isolated from ADSVFs by magnetic cell separation (MACS), and the efficiency of differentiation into chondrocytes was compared with using three methods of pellet method, gel-coating method, and gel-embedding sheet method. Using the CD105-positive cells and the gel-embedding sheet method, aggrecan mRNA was detected about three times higher than pellet and gel-coating methods. The above data suggest that ADSVFs could be differentiated into chondrocyte-like cells in the gel-embedding sheet method and could be useful in regenerative medicine to treat cartilage defects or cartilage degenerative disease. The use of cells sorted by mesenchymal stem cell markers from adipose tissue would gain position in the repair of cartilage tissue.
在大多数发达国家，关节炎疾病的发病率正在迅速上升。关节软骨是关节中最重要的组织，由软骨细胞和包括聚集蛋白聚糖在内的丰富细胞外基质组成，自我修复能力较差。我们研究了小鼠皮下脂肪组织中的干细胞作为再生软骨组织细胞来源的潜力。使用间充质干细胞标志物对脂肪来源的基质血管成分培养细胞（ADSVFs）进行分析，结果显示CD90阳性细胞占93.8%，CD105阳性细胞占68.5%，p75神经营养因子受体（p75NTR，CD271）阳性细胞占36.1%。这些结果表明ADSVFs中存在间充质干细胞标志物阳性的细胞。通过磁性细胞分选（MACS）从ADSVFs中分离出CD105阳性或阴性细胞，并使用沉淀法、凝胶包被法和凝胶包埋片法三种方法比较其向软骨细胞分化的效率。使用CD105阳性细胞和凝胶包埋片法时，检测到的聚集蛋白聚糖mRNA比沉淀法和凝胶包被法高出约三倍。上述数据表明，在凝胶包埋片法中ADSVFs可分化为软骨样细胞，在再生医学中可用于治疗软骨缺损或软骨退行性疾病。利用从脂肪组织中通过间充质干细胞标志物分选的细胞在软骨组织修复中将占据一席之地。
