Comparison of different culture conditions for human mesenchymal stromal cells for clinical stem cell therapy
Comparison of different culture conditions for human mesenchymal stromal cells for clinical stem cell therapy
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DOI：
10.1080/00365510701601681
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发表时间：
2008-01-01
影响因子：
2.1
通讯作者：
Kastrup, J.
中科院分区：
文献类型：
--
作者：
Haack-Sorensen, M.;Friis, T.;Kastrup, J.
Objective.来自成人骨髓的间充质干细胞被认为是治疗心血管疾病的潜在候选者。在临床治疗中实施动物试验结果时，必须在遵循良好生产规范（GMP）的条件下分离和扩增MSC。该研究的目的是首先建立培养条件，遵循GMP质量要求，用于人MSC扩增和分化，用于临床试验，其次将这些MSC与在模拟临床干细胞治疗的动物研究中通常用于MSC培养的四种培养基中培养的MSC进行比较。材料和方法。通过密度梯度离心从BM抽吸物中分离人单核细胞（MNC），并在GMP接受的培养基（EMEA培养基）或四种其他培养基之一中培养。结果流式细胞仪检测结果显示，在EMEA培养基和其他4种培养基中培养的贴壁MSCs均为造血表面标志物CD 45和CD 34阴性，CD 105、CD 73、CD 90、CD 166和CD 13阳性，这5种标志物的联合表达是MSCs的特征。用血管内皮生长因子（VEGF）刺激MSC增加了特征性内皮基因KDR和von Willebrand因子的表达;在蛋白水平上增加了von Willebrand因子和CD 31的表达以及发展毛细血管样结构的能力。结论.我们用符合GMP的培养基建立了用于MSC培养、扩增和分化的培养条件。扩增和分化的MSCs可用于缺血性心脏病患者的自体间充质基质细胞治疗。
Objective. Mesenchymal stromal cells (MSCs) from adult bone marrow (BM) are considered potential candidates for therapeutic neovascularization in cardiovascular disease. When implementing results from animal trials in clinical treatment, it is essential to isolate and expand the MSCs under conditions following good manufacturing practice (GMP). The aims of the study were first to establish culture conditions following GMP quality demands for human MSC expansion and differentiation for use in clinical trials, and second to compare these MSCs with MSCs derived from culture in four media commonly used for MSC cultivation in animal studies simulating clinical stem cell therapy. Material and methods. Human mononuclear cells (MNCs) were isolated from BM aspirates by density gradient centrifugation and cultivated in a GMP-accepted medium (EMEA medium) or in one of four other media. Results. FACS analysis showed that the plastic-adherent MSCs cultured in EMEA medium or in the other four media were identically negative for the haematopoietic surface markers CD45 and CD34 and positive for CD105, CD73, CD90, CD166 and CD13, which in combined expression is characteristic of MSCs. MSC stimulation with vascular endothelial growth factor (VEGF) increased expression of the characteristic endothelial genes KDR and von Willebrand factor; the von Willebrand factor and CD31 at protein level as well as the capacity to develop capillary-like structures. Conclusions. We established culture conditions with a GMP compliant medium for MSC cultivation, expansion and differentiation. The expanded and differentiated MSCs can be used in autologous mesenchymal stromal cell therapy in patients with ischaemic heart disease.