Abstract
Background During the past decade, human stem cells have been the focus of an increased interest due to their potential in clinical applications, as a therapeutic alternative for several diseases. Within this context, human mesenchymal stem cells (hMSCs) have gained special attention due to their immune-modulatory characteristics, as well as in secreting bioactive molecules with anti-inflammatory and regenerative features [1]. In order to face the high demands of hMSCs (from 10 to 10 cells per patient) [2] to be used in therapies, the establishment of robust manufacturing platforms that can ensure the efficient production, purification and formulation of stem cell-based products is still a challenge. Although substantial efforts have been performed on the development of clinical-grade bioprocesses for the expansion of hMSCs in microcarrier-based stirred culture systems, the incorporation of downstream strategies that assure efficient cell-bead separation and consequent hMSC concentration (i.e. volume reduction) and washing is required to deliver safe hMSCs to the clinic [3,4]. Therefore, the main aim of this work was the design of integrated methodologies (filtration and membrane technology approaches) [5] for the robust and clinicalgrade downstream processing of hMSC.
Highlights
During the past decade, human stem cells have been the focus of an increased interest due to their potential in clinical applications, as a therapeutic alternative for several diseases
Materials and methods Cell culture: human mesenchymal stem cells (hMSCs) (STEMCELL TechnologiesTM) were cultivated in IMDM supplemented with 10% of fetal bovine serum (FBS) or in MesenCult®-XF Medium (STEMCELL TechnologiesTM) supplemented with 2 mM
Over the past years, as scale-up platforms for the biomanufacturing of hMSCs become robust enough to yield high cell quantities to support cell-based therapies, culture media supplemented with FBS are becoming less used
Summary
Human stem cells have been the focus of an increased interest due to their potential in clinical applications, as a therapeutic alternative for several diseases. Within this context, human mesenchymal stem cells (hMSCs) have gained special attention due to their immune-modulatory characteristics, as well as in secreting bioactive molecules with anti-inflammatory and regenerative features [1]. In order to face the high demands of hMSCs (from 105 to 109 cells per patient) [2] to be used in therapies, the establishment of robust manufacturing platforms that can ensure the efficient production, purification and formulation of stem cell-based products is still a challenge. Substantial efforts have been performed on the development of clinical-grade bioprocesses for the expansion of hMSCs in microcarrier-based stirred culture systems, the incorporation of downstream strategies that assure efficient cell-bead separation and consequent hMSC concentration (i.e. volume reduction) and washing is required to deliver safe hMSCs to the clinic [3,4].
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