As industrial requirements as well as clinical applications of cell‐based therapeutic treatments require large numbers of cells (Baudequin et al., 2021 Rafiq & Hewitt, 2015 Want et al., 2012), these newly developed thermo‐responsive polymers have been proven to hold clear promises to expand cells with improved cell purity when combined to systems with high surface/volume ratio (S. This technique has been shown not to alter cell physiology, morphology, and immunophenotype of the released cells (Zhang et al., 2015). Thermo‐responsive scaffolds containing poly( N‐isopropylacrylamide) (PNIPAAm) are of great interest, thanks to their specific ability to release cells without physical damage upon temperature changes (Dhamecha et al., 2021 Haq et al., 2017 Kim et al., 2003). This could help moving such approaches towards clinically and industrially relevant processes were cell expansion is needed at very large scale. Furthermore, we validated the scalability potential of both macrobeads production and conjugation with PCL, to produce easily kilograms of thermo‐responsive macrocarriers in a lab environment. By reducing the temperature to 25☌, the MSCs were successfully detached from the PCL‐PNIPAAm beads as observed with fluorescence microscopy. PNIPAAm‐grafted macrobeads allowed mesenchymal stem cells (MSCs) to spread and grow for up to 21 days. The functionalization steps with PNIPAAm were then characterized and confirmed by Fourier transform infrared spectroscopy, scanning electron microscopy, and energy dispersion spectroscopy. 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 size of the beads, all spherical, ranged from 0.5 to 3.7 mm with four bead categories based on production parameters. The effects of PCL concentration on the droplet formation and of flow rate and PVA concentration on the size of the beads were demonstrated.
Following previous studies with PCL‐PNIPAAm carriers, our objectives were to confirm the successful conjugation on homemade macrobeads and to show the advantages of homemade production over commercial beads to control morphological, biological, and fluidization properties.
In this study, polycaprolactone (PCL) macrobeads 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‐isopropylacrylamide) (PNIPAAm) to be used as cell carriers with noninvasive cell detachment properties (thermo‐response).
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