Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells
Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells
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DOI：
10.1002/term.2200
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发表时间：
2017-05-01
影响因子：
3.3
通讯作者：
da Silva, Claudia L.
中科院分区：
文献类型：
--
作者：
de Soure, Antonio M.;Fernandes-Platzgummer, Ana;da Silva, Claudia L.
脐带基质 (UCM) 衍生的间充质干细胞/基质细胞 (MSC) 是再生医学领域有前途的治疗候选者。 UCM MSC 比成体细胞具有优势，因为它们可以通过非侵入性采集程序获得，并表现出更高的增殖能力。然而，临床环境中所需的高细胞剂量使得 UCM MSC 的大规模生产成为强制性的。测试了市售的基于人血小板裂解物的培养补充剂（UltraGRO™，AventaCell BioMedical）（5%（v/v）），以有效分离UCM MSC并在（1）静态条件下使用平面培养系统和（2）在旋转瓶中使用塑料微载体搅拌培养下扩增这些细胞。 11+/-2天后从UCM外植体培养物中分离出MSC样细胞。静态培养五次传代后，UCM MSC 保留了其免疫表型和多谱系分化潜力。与使用先前建立的方案扩增的UCM MSC相比，使用UltraGRO TM 补充培养基在静态条件下培养的UCM MSC扩增得更快。重要的是，使用UltraGRO TM 补充培养基在旋转烧瓶中，UCM MSC 在动态条件下成功地在塑料微载体上扩增。最初 54% 的细胞粘附到珠子上后，UCM MSC 在 5-6 天后扩增超过 13 倍，保持其免疫表型和多谱系分化能力。本文报道了基于人血小板裂解物补充剂的易于扩展的集成培养平台的建立，用于在基于无异种微载体的系统中有效分离和扩增 UCM MSC。该平台代表了在获得更安全且具有临床意义的 MSC 编号以进行临床转化方面的重要进步。版权所有 (C) 2016 约翰·威利父子有限公司
Umbilical cord matrix (UCM)-derived mesenchymal stem/stromal cells (MSCs) are promising therapeutic candidates for regenerative medicine settings. UCM MSCs have advantages over adult cells as these can be obtained through a noninvasive harvesting procedure and display a higher proliferative capacity. However, the high cell doses required in the clinical setting make large-scale manufacturing of UCM MSCs mandatory. A commercially available human platelet lysate-based culture supplement (UltraGRO(TM), AventaCell BioMedical) (5%(v/v)) was tested to effectively isolate UCM MSCs and to expand these cells under (1) static conditions, using planar culture systems and (2) stirred culture using plastic microcarriers in a spinner flask. The MSC-like cells were isolated from UCM explant cultures after 11 +/- 2 days. After five passages in static culture, UCM MSCs retained their immunophenotype and multilineage differentiation potential. The UCM MSCs cultured under static conditions using UltraGRO(TM)-supplemented medium expanded more rapidly compared with UCM MSCs expanded using a previously established protocol. Importantly, UCM MSCs were successfully expanded under dynamic conditions on plastic microcarriers using UltraGRO TM -supplemented medium in spinner flasks. Upon an initial 54% cell adhesion to the beads, UCM MSCs expanded by >13-fold after 5-6 days, maintaining their immunophenotype and multilineage differentiation ability. The present paper reports the establishment of an easily scalable integrated culture platform based on a human platelet lysate supplement for the effective isolation and expansion of UCM MSCs in a xenogeneic-free microcarrier-based system. This platform represents an important advance in obtaining safer and clinically meaningful MSC numbers for clinical translation. Copyright (C) 2016 John Wiley & Sons, Ltd.