Clonal isolation and characterization of bone marrow stromal cells from patients with osteoarthritis
Clonal isolation and characterization of bone marrow stromal cells from patients with osteoarthritis
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DOI：
10.1089/ten.2006.0180
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发表时间：
2007-04-01
影响因子：
--
通讯作者：
Xiao, Yin
中科院分区：
文献类型：
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作者：
Mareddy, Shobha;Crawford, Ross;Xiao, Yin
对受损肌肉骨骼组织治疗策略的需求不断增长，特别是随着越来越多的老年人患有骨关节炎（OA）等骨骼系统退行性疾病。由于多能细胞的耗尽与退行性关节疾病有关，因此已经提出了用于组织再生，特别是用于软骨修复的基于细胞的疗法。为探讨从骨关节炎患者骨髓中分离、扩增多能间充质干细胞（mesenchymal stem cells，MSCs）的可能性，采用有限稀释法从3例骨关节炎患者的骨髓中分离、扩增MSCs，建立单细胞克隆培养体系。建立了14个克隆用于随后的表征。细胞倍增时间有很大的差异，达到20个群体倍增所需的时间从37天到100多天不等。将这些无性系分为快速生长无性系和缓慢生长无性系。除了一个快速生长的干细胞克隆外，所有克隆都是三能的。然而，生长缓慢的克隆表现出有限的分化潜力和形态变化与细胞衰老与培养时间延长。流式细胞仪分析表明，强烈需要调查新的细胞表面特征标记的BMSC，因为有没有明显的差异，在表达所选择的特征性BMSC细胞表面标记CD 29，CD 44，CD 90，CD 105，和CD 166之间的快速增长和缓慢增长的克隆。这项研究表明，存在一个快速增长的多潜能MSC人口从骨髓样本的OA患者。因此，尽管这些患者的干细胞室较小，但我们在这里证明它们仍然可以产生潜在的治疗有用的同源MSC来源。
The demand for treatment strategies for damaged musculoskeletal tissue is continuously growing, especially with the increasing number of older people with degenerative diseases of the skeletal system such as osteoarthritis (OA). Because depletion of multipotent cells has been implicated in degenerative joint diseases, cell-based therapies have been proposed for tissue regeneration, especially for cartilage repair. The aim of the present study is to focus on the possibility of deriving and expanding multipotential mesenchymal stem cells (MSCs) from bone marrow samples of patients with OA by characterizing MSCs at the single cell level.Single-cell clonal cultures were established in 96-well plates by limiting dilution of bone marrow stromal cells (BMSCs) from three patients with OA. Fourteen clones were established for subsequent characterization. There was a wide variation in cell doubling times, with the time taken to reach 20 population doublings ranging from 37 days to more than 100 days. The clones were grouped into fast-growing and slow-growing clones. All except one of the fast-growing stem cell clones were tripotential. However, the slow-growing clones showed limited differentiation potential and morphological changes associated with cellular senescence with extended duration in culture. Flow cytometric analysis indicated a strong need to investigate for novel cell-surface characteristic markers of BMSCs because there was no obvious difference in the expression of the selected characteristic BMSC cell surface markers CD29, CD44, CD90, CD105, and CD166 between fast-growing and slow-growing clones. This study has demonstrated the existence of a fast-growing multipotential MSC population from bone marrow samples of patients with OA. Therefore, despite a supposedly smaller stem cell compartment in these patients, we demonstrate here that they can still yield a potentially therapeutically useful source of syngeneic MSCs.