Sakaguchi Yusuke, Sekiya Ichiro, Yagishita Kazuyoshi, Muneta Takeshi
Tokyo Medical and Dental University, Japan.
Arthritis Rheum. 2005 Aug;52(8):2521-9. doi: 10.1002/art.21212.
To compare the properties of human mesenchymal stem cells (MSCs) isolated from bone marrow, synovium, periosteum, skeletal muscle, and adipose tissue.
Human mesenchymal tissues were obtained from 8 donors during knee surgery for ligament injury. After collagenase digestion or gradient-density separation, nucleated cells were plated at an appropriate density for expansion at the maximum rate without colony-to-colony contact. Yield, expandability, differentiation potential, and epitope profile were compared among MSCs from the 5 different tissue sources.
Colony number per 10(3) nucleated cells was lower, and cell number per colony was higher, in bone marrow than in other mesenchymal tissues. When the cells were replated at low density every 14 days, bone marrow-, synovium-, and periosteum-derived cells retained their proliferation ability even at passage 10. In chondrogenesis studies in which the cells were pelleted and cultured in vitro, pellets from bone marrow-, synovium-, and periosteum-derived cells were shown to be larger and stained more extensively for cartilage matrix. Synovium-derived cells, in particular, had the greatest ability for chondrogenesis. In adipogenesis experiments, the frequency of oil red O-positive colonies was highest in synovium- and adipose tissue-derived cells. In studies of osteogenesis, the rate of alizarin red-positive colonies was highest in bone marrow-, synovium-, and periosteum-derived cells. For epitope profiling, 15 surface antigens were measured. Most appeared to have similar epitope profiles irrespective of cell source.
Our findings indicate that there are significant differences in MSC properties according to tissue source, beyond donor and experimental variation. Superiority of synovium as a potential source of MSCs for clinical applications was demonstrated.
比较从骨髓、滑膜、骨膜、骨骼肌和脂肪组织中分离出的人间充质干细胞(MSC)的特性。
在膝关节韧带损伤手术中从8名供体获取人间充质组织。经胶原酶消化或梯度密度分离后,将有核细胞以适当密度接种,以便在不发生集落间接触的情况下以最大速率扩增。比较了来自5种不同组织来源的MSC的产量、扩增能力、分化潜能和表位谱。
每10³个有核细胞中的集落数,骨髓中的比其他间充质组织中的少,而每个集落中的细胞数则较多。当每14天将细胞以低密度重新接种时,来自骨髓、滑膜和骨膜的细胞即使在传代10次时仍保留其增殖能力。在细胞制成小球并进行体外培养的软骨形成研究中,来自骨髓、滑膜和骨膜的细胞形成的小球更大,软骨基质染色更广泛。特别是滑膜来源的细胞具有最强的软骨形成能力。在脂肪形成实验中,油红O阳性集落的频率在滑膜和脂肪组织来源的细胞中最高。在成骨研究中,茜素红阳性集落的比率在骨髓、滑膜和骨膜来源的细胞中最高。对于表位分析,检测了15种表面抗原。无论细胞来源如何,大多数表面抗原似乎具有相似的表位谱。
我们的研究结果表明,除了供体和实验差异外,根据组织来源,MSC特性存在显著差异。滑膜作为临床应用中潜在的MSC来源的优越性得到了证实。