Institute of Clinical and Experimental Transfusion Medicine (IKET), University Hospital Tübingen, Otfried-Müller-Strasse 4/1, Tübingen, D-72076, Germany.
BMC Med. 2013 Jun 11;11:146. doi: 10.1186/1741-7015-11-146.
Mesenchymal stromal cells (MSCs) are attractive for cell-based therapies ranging from regenerative medicine and tissue engineering to immunomodulation. However, clinical efficacy is variable and it is unclear how the phenotypes defining bone marrow (BM)-derived MSCs as well as donor characteristics affect their functional properties.
BM-MSCs were isolated from 53 (25 female, 28 male; age: 13 to 80 years) donors and analyzed by: (1) phenotype using flow cytometry and cell size measurement; (2) in vitro growth kinetics using population doubling time; (3) colony formation capacity and telomerase activity; and (4) function by in vitro differentiation capacity, suppression of T cell proliferation, cytokines and trophic factors secretion, and hormone and growth factor receptor expression. Additionally, expression of Oct4, Nanog, Prdm14 and SOX2 mRNA was compared to pluripotent stem cells.
BM-MSCs from younger donors showed increased expression of MCAM, VCAM-1, ALCAM, PDGFRβ, PDL-1, Thy1 and CD71, and led to lower IL-6 production when co-cultured with activated T cells. Female BM-MSCs showed increased expression of IFN-γR1 and IL-6β, and were more potent in T cell proliferation suppression. High-clonogenic BM-MSCs were smaller, divided more rapidly and were more frequent in BM-MSC preparations from younger female donors. CD10, β1integrin, HCAM, CD71, VCAM-1, IFN-γR1, MCAM, ALCAM, LNGFR and HLA ABC were correlated to BM-MSC preparations with high clonogenic potential and expression of IFN-γR1, MCAM and HLA ABC was associated with rapid growth of BM-MSCs. The mesodermal differentiation capacity of BM-MSCs was unaffected by donor age or gender but was affected by phenotype (CD10, IFN-γR1, GD2). BM-MSCs from female and male donors expressed androgen receptor and FGFR3, and secreted VEGF-A, HGF, LIF, Angiopoietin-1, basic fibroblast growth factor (bFGF) and NGFB. HGF secretion correlated negatively to the expression of CD71, CD140b and Galectin 1. The expression of Oct4, Nanog and Prdm14 mRNA in BM-MSCs was much lower compared to pluripotent stem cells and was not related to donor age or gender. Prdm14 mRNA expression correlated positively to the clonogenic potential of BM-MSCs.
By identifying donor-related effects and assigning phenotypes of BM-MSC preparations to functional properties, we provide useful tools for assay development and production for clinical applications of BM-MSC preparations.
间充质基质细胞（MSCs）在细胞治疗中具有吸引力，范围从再生医学和组织工程到免疫调节。然而，临床疗效是可变的，目前尚不清楚定义骨髓（BM）来源的 MSCs 的表型以及供体特征如何影响其功能特性。
从 53 名（25 名女性，28 名男性；年龄：13 至 80 岁）供体中分离 BM-MSCs，并通过以下方法进行分析：（1）使用流式细胞术和细胞大小测量进行表型分析；（2）通过群体倍增时间分析体外生长动力学；（3）集落形成能力和端粒酶活性；（4）通过体外分化能力、抑制 T 细胞增殖、细胞因子和营养因子分泌以及激素和生长因子受体表达来评估功能。此外，将 Oct4、Nanog、Prdm14 和 SOX2 mRNA 的表达与多能干细胞进行了比较。
来自年轻供体的 BM-MSCs 表现出 MCAM、VCAM-1、ALCAM、PDGFRβ、PDL-1、Thy1 和 CD71 的表达增加，并且在与激活的 T 细胞共培养时产生的 IL-6 减少。女性 BM-MSCs 表现出 IFN-γR1 和 IL-6β 的表达增加，并且在 T 细胞增殖抑制方面更为有效。高克隆形成能力的 BM-MSCs 更小，分裂更快，并且在来自年轻女性供体的 BM-MSC 制剂中更为常见。CD10、β1 整联蛋白、HCAM、CD71、VCAM-1、IFN-γR1、MCAM、ALCAM、LNGFR 和 HLA ABC 与具有高克隆形成潜力的 BM-MSC 制剂相关，IFN-γR1、MCAM 和 HLA ABC 的表达与 BM-MSCs 的快速生长相关。BM-MSCs 的中胚层分化能力不受供体年龄或性别影响，但受表型（CD10、IFN-γR1、GD2）影响。来自女性和男性供体的 BM-MSCs 表达雄激素受体和 FGFR3，并分泌 VEGF-A、HGF、LIF、血管生成素-1、碱性成纤维细胞生长因子（bFGF）和 NGFB。HGF 的分泌与 CD71、CD140b 和半乳糖凝集素 1 的表达呈负相关。BM-MSCs 中 Oct4、Nanog 和 Prdm14 mRNA 的表达远低于多能干细胞，与供体年龄或性别无关。Prdm14 mRNA 的表达与 BM-MSCs 的克隆形成潜力呈正相关。
通过鉴定与供体相关的影响，并将 BM-MSC 制剂的表型分配给功能特性，我们为 BM-MSC 制剂的检测开发和生产提供了有用的工具，可用于临床应用。
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