源自滑膜的间充质基质细胞的表面表征及软骨形成分化
Surface characterization and chondrogenic differentiation of mesenchymal stromal cells derived from synovium.
作者信息
Jo C H, Ahn H J, Kim H J, Seong S C, Lee M C
机构信息
Department of Orthopedic Surgery, Seoul National University Boramae Hospital, Dongjak-gu, Seoul, Korea.
出版信息
Cytotherapy. 2007;9(4):316-27. doi: 10.1080/14653240701291620.
BACKGROUND
Synovium is the only tissue that can produce hyaline cartilage in benign conditions, such as synovial chondromatosis and osteoarthritis, suggesting potential advantages in chondrogenesis using mesenchymal stromal cells. We performed surface characterization of cells isolated from the synovium of patients with osteoarthritis after different passages and induced chondrogenic differentiation.
METHODS
Using cells obtained from synovium, colony-forming unit fibroblast assay and characterization of cell-surface markers by flow cytometry using 22 different Ab at different passages were performed. Cells were cultured under chondrogenic conditions and evaluated grossly, histologically, immunohistochemically and by [(35)S]sulfate incorporation and reverse transcription-PCR.
RESULTS
The positive cell-surface markers of immediately isolated cells were CD10, CD13, CD14, CD34, CD44, CD45, CD49a, CD62e, CD73 and HLA-DR. After the first passage (P), CD14, CD34, CD45, CD62e and HLA-DR disappeared, whereas CD105 and CD166 appeared and CD10, CD13, CD44, CD49a and CD73 showed increased expression levels. The surface marker expression level did not vary much after P1 through to P8. The chondrogenic differentiation potential of cells from the synovium was confirmed using various evaluation methods.
DISCUSSION
We have demonstrated that cells from synovium contain a mesenchymal stromal cell population capable of chondrogenic differentiation, which seems to increase with passage under our culture conditions. The cell-surface markers were found to change remarkably after the first passage and then remained stable. The results of this study may be helpful for sorting mesenchymal stromal cells from heterogeneous synovial cells for future studies.
背景
滑膜是在诸如滑膜软骨瘤病和骨关节炎等良性病症中唯一能够产生透明软骨的组织,这表明在利用间充质基质细胞进行软骨形成方面具有潜在优势。我们对从骨关节炎患者滑膜中分离出的细胞在不同传代后进行了表面特征分析,并诱导其向软骨细胞分化。
方法
使用从滑膜获得的细胞,进行集落形成单位成纤维细胞测定,并通过流式细胞术使用22种不同抗体在不同传代时对细胞表面标志物进行特征分析。将细胞在软骨形成条件下培养,并通过大体观察、组织学、免疫组织化学以及[³⁵S]硫酸盐掺入和逆转录-聚合酶链反应进行评估。
结果
刚分离出的细胞的阳性细胞表面标志物为CD10、CD13、CD14、CD34、CD44、CD45、CD49a、CD62e、CD73和HLA-DR。首次传代(P1)后,CD14、CD34、CD45、CD62e和HLA-DR消失,而CD105和CD166出现,且CD10、CD13、CD44、CD49a和CD73表达水平升高。从P1到P8,表面标志物表达水平变化不大。使用多种评估方法证实了滑膜来源细胞的软骨形成分化潜能。
讨论
我们已经证明,滑膜来源的细胞包含能够进行软骨形成分化的间充质基质细胞群体,在我们的培养条件下,这种能力似乎随着传代而增强。发现细胞表面标志物在首次传代后发生显著变化,然后保持稳定。本研究结果可能有助于从异质滑膜细胞中筛选间充质基质细胞以供未来研究使用。