Growth and Functional Harvesting of Human Mesenchymal Stromal Cells Cultured on a Microcarrier-Based System
Growth and Functional Harvesting of Human Mesenchymal Stromal Cells Cultured on a Microcarrier-Based System
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DOI：
10.1002/btpr.1886
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发表时间：
2014-07-01
影响因子：
2.9
通讯作者：
Swiech, Kamilla
中科院分区：
文献类型：
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作者：
Caruso, Samia R.;Orellana, Maristela D.;Swiech, Kamilla
人骨髓间充质干细胞(HMSCs)在组织工程和细胞治疗方面具有广阔的应用前景。由于hMSCs在组织中的利用率低以及输注所需的高剂量hMSCs，因此需要可扩展的、经济有效的体外细胞扩增技术来生产MSCs，同时保持其功能、免疫表型和细胞遗传学特性。基于微载体的培养系统是传统培养系统的良好替代方案。本研究的目的是开发一种可扩展的人骨髓间充质干细胞(HBM-MSCs)在微载体上扩增的生物过程，以优化生长和功能采集。总体而言，实验结果证明了利用微载体技术扩增HBM-MSCs的可行性。第7天的最大细胞浓度(n=5)接近4.82+/-1.18×10(5)cell mL(-1)，是接种细胞数量的3.9倍。培养结束时，细胞回收率为87.2%(存活率为95%)。细胞代谢分析表明，在培养过程中没有消耗葡萄糖和谷氨酰胺等重要营养物质，在抑制浓度下也没有形成乳酸和氨的副产物。扩增后恢复的细胞保持其免疫表型和功能特征。这些结果代表着朝着实施符合GMP的大规模生产系统的方向迈出了重要的一步，hMSCs用于细胞治疗。(C)2014年美国化学工程师学会
Human mesenchymal stromal cells (hMSCs) cells are attractive for applications in tissue engineering and cell therapy. Because of the low availability of hMSCs in tissues and the high doses of hMSCs necessary for infusion, scalable and cost-effective technologies for in vitro cell expansion are needed to produce MSCs while maintaining their functional, immunophenotypic and cytogenetic characteristics. Microcarrier-based culture systems are a good alternative to traditional systems for hMSC expansion. The aim of the present study was to develop a scalable bioprocess for the expansion of human bone marrow mesenchymal stromal cells (hBM-MSCs) on microcarriers to optimize growth and functional harvesting. In general, the results obtained demonstrated the feasibility of expanding hBM-MSCs using microcarrier technology. The maximum cell concentration (n = 5) was similar to 4.82 +/- 1.18 x 10(5) cell mL(-1) at day 7, representing a 3.9-fold increase relative to the amount of inoculated cells. At the end of culture, 87.2% of the cells could be harvested (viability = 95%). Cell metabolism analysis revealed that there was no depletion of important nutrients such as glucose and glutamine during culture, and neither lactate nor ammonia byproducts were formed at inhibitory concentrations. The cells that were recovered after the expansion retained their immunophenotypic and functional characteristics. These results represent an important step toward the implementation of a GMP-compliant large-scale production system for hMSCs for cellular therapy. (C) 2014 American Institute of Chemical Engineers