Umbilical cord derived mesenchymal stromal cells in microcarrier based industrial scale culture sustain the immune regulatory functions

Abstract

Mesenchymal stromal cells (MSCs) have been isolated from numerous sources and are potentially therapeutic against various diseases. Umbilical cord‐derived MSCs (UC‐MSCs) are considered superior to other tissue‐derived MSCs since they have a higher proliferation rate and can be procured using less invasive surgical procedures. However, it has been recently reported that 2D culture systems, using conventional cell culture flasks, limit the mass production of MSCs for cell therapy. Therefore, the development of alternative technologies, including microcarrier‐based cell culture in bioreactors, is required for the large‐scale production and industrialization of MSC therapy. In this study, we aimed to optimize the culture conditions for UC‐MSCs by using a good manufacturing practice (GMP)‐compatible serum‐free medium, developed in‐house, and a small‐scale (30 mL) bioreactor, which was later scaled up to 500 mL. UC‐MSCs cultured in microcarrier‐based bioreactors (MC‐UC‐MSCs) showed characteristics equivalent to those cultured statically in conventional cell culture flasks (ST‐UC‐MSCs), fulfilling the minimum International Society for Cellular Therapy criteria for MSCs. Additionally, we report, for the first time, the equivalent therapeutic effect of MC‐UC‐MSCs and ST‐UC‐MSCs in immunodeficient mice (graft‐versus‐host disease model). Lastly, we developed a semi‐automated cell dispensing system, without bag‐to‐bag variation in the filled volume or cell concentration. In summary, our results show that the combination of our GMP‐compatible serum‐free and microcarrier‐based culture systems is suitable for the mass production of MSCs at an industrial scale. Further improvements in this microcarrier‐based cell culture system can contribute to lowering the cost of therapy and satisfying several unmet medical needs.

A Scalable System for Generation of Mesenchymal Stem Cells Derived from Induced Pluripotent Cells Employing Bioreactors and Degradable Microcarriers

“…When ihMSCs were co-incubated with murine splenocytes that were pre-activated with LPS, a dose-dependent reduction in the secretion of inflammatory cytokines was observed, indicating that the bioreactor-generated ihMSCs have the theoretical capacity to blunt early-stage inflammatory responses triggered by the engagement of toll-like receptor 4 (TLR4) 70 and therefore may have the capacity to modulate inflammatory responses in vivo in a similar manner to tissue-derived hMSCs. 71,72 Bioreactor-expanded sources of tissue-derived hMSCs have been reported to blunt inflammatory responses in in vitro assays, 73,74 and the immunomodulatory capacity of iPSC-derived hMSCs has been demonstrated, 25 but to our knowledge, this is the first time immunomodulatory ihMSCs have been generated using a scalable VWB platform.…”

A large‐scale production of mesenchymal stem cells and their exosomes for an efficient treatment against lung inflammation

“…For example, the volume of the bioreactor, the cell density, the geometry of the cell colonization on microcarriers, the cell collection time, and the feeding regimen are suggested as the possible impacting factors. [ 13,15,39 ]…”

“…For example, the volume of the bioreactor, the cell density, the geometry of the cell colonization on microcarriers, the cell collection time, and the feeding regimen are suggested as the possible impacting factors. [13,15,39] Additionally, exosomes secreted by MSCs were revealed to have various anti-inflammatory relevant miRNAs, mRNAs, and cytokines, which were responsible for the inflammatory regulation. [40,41] For example, miR-23a-3p and miR-182-5p delivered by MSC-derived exosomes are believed to participate in the regulation of lung inflammation.…”

“…Studies have demonstrated that the culture method during the process of large-scale production may affect the quality and biofunction of stem cells. [12,13] For example, large-scale adherent cell culture may have a negative effect on the proliferation and differentiation of stem cells. [14] To overcome this obstacle, the recently emerged 3D culture technology was introduced into the large-scale production of stem cells, showing the advantages of improving the proliferation ability and improving the batch-to-batch cell stability.…”

Evaluation of Polymeric Particles for Modular Tissue Cultures in Developmental Engineering

“…The advantages of these microcarrier-based expansion systems include: (i) ease of scale up [ 29 , 35 , 36 , 37 ], (ii) ability to precisely control cell growth conditions within large-scale bioreactors [ 38 , 39 ], (iii) reduction in bioreactor volumes and the floor spaces required for given-sized manufacturing operations [ 40 ], (iv) reduction in technician labor [ 37 , 39 ], and (v) more natural culture environments for cell proliferation and differentiation [ 38 , 41 ]. All these benefits have enabled the microcarrier-based culture technology to be generally employed for industrial production of mammalian cells [ 37 , 42 ], protein-based therapeutics [ 29 ], and for research purposes [ 35 ]. Furthermore, it potentially provides a straightforward scale-up strategy for modular tissue cultures in DE.…”