Characterization of bone marrow derived mesenchymal stem cells in suspension.
Characterization of bone marrow derived mesenchymal stem cells in suspension.
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DOI：
10.1186/scrt131
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发表时间：
2012-10-19
影响因子：
7.5
通讯作者：
Shi S
中科院分区：
文献类型：
--
作者：
Akiyama K;You YO;Yamaza T;Chen C;Tang L;Jin Y;Chen XD;Gronthos S;Shi S
骨髓间充质干细胞（BMMSC）是一种异质的出生后前体细胞群，具有粘附于培养皿产生集落形成单位成纤维细胞（CFU-F）的能力。在这里，我们确定了一个新的BMMSCs的子集，不能坚持塑料培养皿，并保持在培养悬浮液（S-BMMSCs）。为了捕获S-BMMSC，我们使用BMMSC产生的细胞外基质（ECM）包被的培养皿。采用流式细胞术、诱导分化、western blot和体内移植等方法对分离的S-BMMSCs进行体外干细胞分析，评价S-BMMSCs的成骨能力。此外，我们进行了系统性S-BMMSCs移植治疗系统性红斑狼疮（SLE）样MRL/lpr小鼠。S-BMMSC能够粘附于ECM包被的培养皿，并显示出与造血细胞不同的间充质干细胞特征，如通过CD 73或Oct-4与CD 34的共表达所证明的，在ECM上形成单集落簇，并且不能分化成造血细胞谱系。此外，我们发现培养扩增的S-BMMSCs在体外表现出显著增加的免疫调节能力，并通过高NO产生来重新平衡调节性T细胞（TCLs）和辅助性T细胞17（Th 17），从而有效治疗SLE样MRL/lpr小鼠。这些数据表明，它是可行的，以改善免疫治疗，通过确定一个新的子集BMMSCs。
Bone marrow mesenchymal stem cells (BMMSCs) are a heterogeneous population of postnatal precursor cells with the capacity of adhering to culture dishes generating colony-forming unit-fibroblasts (CFU-F). Here we identify a new subset of BMMSCs that fail to adhere to plastic culture dishes and remain in culture suspension (S-BMMSCs). To catch S-BMMSCs, we used BMMSCs-produced extracellular cell matrix (ECM)-coated dishes. Isolated S-BMMSCs were analyzed by in vitro stem cell analysis approaches, including flow cytometry, inductive multiple differentiation, western blot and in vivo implantation to assess the bone regeneration ability of S-BMMSCs. Furthermore, we performed systemic S-BMMSCs transplantation to treat systemic lupus erythematosus (SLE)-like MRL/lpr mice. S-BMMSCs are capable of adhering to ECM-coated dishes and showing mesenchymal stem cell characteristics with distinction from hematopoietic cells as evidenced by co-expression of CD73 or Oct-4 with CD34, forming a single colony cluster on ECM, and failure to differentiate into hematopoietic cell lineage. Moreover, we found that culture-expanded S-BMMSCs exhibited significantly increased immunomodulatory capacities in vitro and an efficacious treatment for SLE-like MRL/lpr mice by rebalancing regulatory T cells (Tregs) and T helper 17 cells (Th17) through high NO production. These data suggest that it is feasible to improve immunotherapy by identifying a new subset BMMSCs.
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