Development and validation of broad-spectrum magnetic particle labelling processes for cell therapy manufacturing

Abstract

BackgroundStem cells are increasingly seen as a solution for many health challenges for an ageing population. However, their potential benefits in the clinic are currently curtailed by technical challenges such as high cell dose requirements and point of care delivery, which pose sourcing and logistics challenges. Cell manufacturing solutions are currently in development to address the supply issue, and ancillary technologies such as nanoparticle-based labelling are being developed to improve stem cell delivery and enable post-treatment follow-up.MethodsThe application of magnetic particle (MP) labelling to potentially scalable cell manufacturing processes was investigated in a range of therapeutically relevant cells, including mesenchymal stromal cells (MSC), cardiomyocytes (CMC) and neural progenitor cells (ReN). The efficiency and the biological effect of particle labelling were analysed using fluorescent imaging and cellular assays.ResultsFlow cytometry and fluorescent microscopy confirmed efficient labelling of monolayer cultures. Viability was shown to be retained post labelling for all three cell types. MSC and CMC demonstrated higher tolerance to MP doses up to 100× the standard concentration. This approach was also successful for MP labelling of suspension cultures, demonstrating efficient MP uptake within 3 h, while cell viability was unaffected by this suspension labelling process. Furthermore, a procedure to enable the storing of MP-labelled cell populations to facilitate cold chain transport to the site of clinical use was investigated. When MP-labelled cells were stored in hypothermic conditions using HypoThermosol solution for 24 h, cell viability and differentiation potential were retained post storage for ReN, MSC and beating CMC.ConclusionsOur results show that a generic MP labelling strategy was successfully developed for a range of clinically relevant cell populations, in both monolayer and suspension cultures. MP-labelled cell populations were able to undergo transient low-temperature storage whilst maintaining functional capacity in vitro. These results suggest that this MP labelling approach can be integrated into cell manufacturing and cold chain transport processes required for future cell therapy approaches.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0968-0) contains supplementary material, which is available to authorized users.

In Vitro Innovation of Tendon Tissue Engineering Strategies

“…Moreover, magnetic nanoparticles can be attached to the cell membrane, and specific receptors can be activated using Alternating Magnetic Fields (AMFs) [298,305,306]. As a consequence of these simulations, a mechanotransduction event is obtained, evidencing the ability of cells to respond to mechanical stimuli using biochemical signals [307] Specific receptors on the cell membrane can be tagged with magnetic nanoparticles and then mechanoactivated with remote magnetic fields [308,309]. This approach was explored to induce the tenogenic differentiation of hASCs.…”

Magnetically Assisted Control of Stem Cells Applied in 2D, 3D and In Situ Models of Cell Migration

“…Building on this concept of magnetic targeting, the use of iron oxide particles to label cells intracellularly has also been proposed as a way to confer them magneto-responsiveness [2,3,33]. Iron oxide particle internalisation for cell labelling was shown to be safe in a range of cell types using in vitro and in vivo models [9,34]. This represents a promising solution to develop better cell targeting and improve stem cell localisation and engraftment for tissue engineering.…”

“…Results presented here highlight the application of magnetic forces to control MSCs in culture using experimental models of cell recruitment, immobilisation and patterning. The design of magnetic arrays has been proposed as a tissue assembly and culture engineering tool [35,36], which could easily be combined with the labelling approach presented here, exploiting the efficient spontaneously internalisation of silica-shell MPIO by MSCs and other cell types [8,9]. This method may be easier and more readily accessible than other multicell assembling methods such as optical tweezers, which depend on highly advanced technology [37].…”

“…Iron oxide nanoparticles have been developed as contrast agents employed for advanced MRI imaging in vivo [4,5,6], and more recently as theranostics mediators, enabling the application of targeted hyperthermia for tumour treatment [7]. Their cytocompatibility and capacity for efficient cellular uptake also made them applicable to in vitro studies, facilitated by the availability of commercial fluorescently tagged particles compatible with confocal and advanced super-resolution imaging [8,9,10]. Efficient spontaneous uptake of MPIO has been documented in a range of cell types including human stem cells, and was recently shown to be compatible with nanoparticle-based intracellular sensing [10].…”

Magnetic biomaterials and nano-instructive tools as mediators of tendon mechanotransduction

“…An alternative approach is to magnetically tag specic receptors on the cells with magnetic nanoparticles 93,94 which have been functionalized with specic receptor targets which can be mechano-activated via remote magnetic elds. Magnetic mechano-activation remotely delivers mechanical stimuli directly to cells which are transmitted through activation of mechanically sensitive receptors available on the cell membrane.…”