Production of Mesenchymal Progenitor Cell-Derived Extracellular Vesicles in Suspension Bioreactors for Use in Articular Cartilage Repair

Abstract

Mesenchymal progenitor cells (MPCs) have shown promise initiating articular cartilage repair, with benefits largely attributed to the trophic factors they secrete. These factors can be found in the conditioned medium (CM) collected from cell cultures, and it is believed that extracellular vesicles (EVs) within this CM are at least partially responsible for MPC therapeutic efficacy. This study aimed to examine the functionality of the EV fraction of CM compared to whole CM obtained from human adipose-derived MPCs in an in vivo murine cartilage defect model. Mice treated with whole CM or the EV fraction demonstrated an enhanced cartilage repair score and type II collagen deposition at the injury site compared to saline controls. We then developed a scalable bioprocess using stirred suspension bioreactors (SSBs) to generate clinically relevant quantities of MPC-EVs. Whereas static monolayer culture systems are simple to use and readily accessible, SSBs offer increased scalability and a more homogenous environment due to constant mixing. This study evaluated the biochemical and functional properties of MPCs and their EV fractions generated in static culture versus SSBs. Functionality was assessed using in vitro MPC chondrogenesis as an outcome measure. SSBs supported increased MPC expression of cartilage-specific genes, and EV fractions derived from both static and SSB culture systems upregulated type II collagen production by MPCs. These results suggest that SSBs are an effective platform for the generation of MPC-derived EVs with the potential to induce cartilage repair.

Culture of Hoffa fat pad mesenchymal stem/stromal cells on microcarrier suspension in vertical wheel bioreactor for extracellular vesicle production

“…Production of MSC EVs for therapeutic applications is a growing field also in the context of bone and cartilage diseases [ 14 , 15 , 19 , 22 , 27 ]. Different 3D culture approaches are investigated including stirred tank bioreactors [ 32 ] and vertical wheel bioreactors [ 6 ], however, there are still standardisations to be developed and problems to be solved. One of them is the decrease in cell number during EV collection observed in this study (Fig.…”

The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases

“…Moreover, stirred suspension bioreactors (SSBs) have been efficiently utilized to scale up MSCs manufacture. These techniques are partly simple to perform, propose scalability benefits, supply an excellent capacity for cell development, can help adherent cell development with the addition of microcarriers or accumulations, and allow a homogenous culture microenvironment because of continuous stirring [ 70 ] (Fig. 5 ).…”

Bioengineering extracellular vesicles: smart nanomaterials for bone regeneration

“…Microcarriers have been extensively employed for the manufacture of clinical-grade cells in conjugation with bioreactors systems [ 174 , 175 ]. Due to their extensive use in cell manufacture, increasing studies have harnessed microcarrier-based bioreactor systems for EV production, with promising results observed [ 154 , 176 ]. Three classes of microcarriers are commonly used for cell manufacturing, which are non-porous, microporous and macroporous in structure.…”