Validation and scalability of homemade polycaprolactone macro-beads grafted with thermo-responsive poly (N-isopropyl acrylamide) for mesenchymal stem cell expansion and harvesting.
Nguyen LTB., Baudequin T., Cui Z., Ye H.
In this study, polycaprolactone (PCL) macro-beads were prepared by an oil-in-water (o/w) emulsion solvent evaporation method with poly (vinyl alcohol) (PVA) as an emulsifier and conjugated to poly (N-isopropyl acrylamide) (PNIPAAm) to be used as cell carriers with non-invasive cell detachment properties (thermo-response). Following previous studies with PCL-PNIPAAm carriers, our objectives were to confirm the successful conjugation on homemade macro-beads and to show the advantages of homemade production over commercial beads to control morphological, biological and fluidization properties. The effects of PCL concentration on the droplet formation and of flow rate and PVA concentration on the size of the beads were demonstrated. The size of the beads, all spherical, ranged from 0.5 mm to 3.7 mm with four bead categories based on production parameters. The morphology and size of the beads were observed by scanning electron microscopy to show surface roughness enhancing cell attachment and proliferation compared to commercial beads. The functionalization steps with PNIPAAm were then characterized and confirmed by Fourier transform infrared spectroscopy (FTIR), SEM and Energy Dispersion Spectroscopy (EDS). PNIPAAm-grafted macro-beads allowed mesenchymal stem cells (MSC) to spread and grow for up to 21 days. By reducing the temperature to 25 °C, the MSCs were successfully detached from the PCL-PNIPAAm beads as observed with fluorescence microscopy. Furthermore, we validated the scalability potential of both macro-beads production and conjugation with PCL, in order to produce easily kilograms of thermo-responsive macro-carriers in a lab environment. This could help moving such approaches towards clinically and industrially relevant processes were cell expansion is needed at very large scale. This article is protected by copyright. All rights reserved.