Monitoring MSCs during expansion in a stirred tank bioreactor

      Human mesenchymal stromal/stem cells (hMSCs) are an attractive target for clinical study as therapeutic agents. However, multilayer flatbed culture expansion paradigms are cumbersome, time consuming and typically limited in the ability to monitor cell characteristics during the growth process. We described a new expansion paradigm that uses a scalable, single use, stirred tank bioreactor with microcarrier scaffold for hMSC expansion. In addition to ease of handling improvements and lower medium volume requirements, the bioreactor system enables sampling of the hMSCs at any time during the expansion campaign. The cells can then be assessed for a variety of markers that denote desired identity and purity, and for expected cell health status. Beyond the obvious cell viability assessment, apoptosis and cell cycle provide a toolbox of assays for cell health. Changes in the protein markers or cell health status can be used during process development to identify optimal conditions or as a process quality assessment following expansion. In this work, we demonstrate that the combination of cell health assays and protein markers for identity and purity are key assessment tools to support and characterize the expansion of hMSCs in a scalable bioreactor system.
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