Efficient manufacturing of therapeutic mesenchymal stromal cells with the use of the Quantum Cell Expansion System

Abstract

Background The use of bone marrow-derived mesenchymal stromal cells (MSCs) as a cellular therapy for various diseases, such as graft-versus-host-disease, diabetes, ischemic cardiomyopathy, and Crohn's disease has produced promising results in early-phase clinical trials. However, for widespread application and use in later phase studies, manufacture of these cells needs to be cost effective, safe, and reproducible. Current methods of manufacturing in flasks or cell factories are labor-intensive, involve a large number of open procedures, and require prolonged culture times. Methods We evaluated the Quantum Cell Expansion system for the expansion of large numbers of MSCs from unprocessed bone marrow in a functionally closed system and compared the results to a flask-based method currently in clinical trials. Results After only two passages, we were able to expand a mean of 6.6×108 MSCs from 25 mL of bone marrow reproducibly. The mean expansion time was 21 days, and cells obtained were able to differentiate into all three lineages: chondrocytes, osteoblasts, and adipocytes. The Quantum was able to generate the target cell number of 2.0×108 cells in an average of 9-fewer days and in half the number of passages required during flask-based expansion. We estimated the Quantum would involve 133 open procedures versus 54,400 in flasks when manufacturing for a clinical trial. Quantum-expanded MSCs infused into an ischemic stroke rat model were therapeutically active. Discussion The Quantum is a novel method of generating high numbers of MSCs in less time and at lower passages when compared to flasks. In the Quantum, the risk of contamination is substantially reduced due to the substantial decrease in open procedures.

Clinical grade expansion of MSCs

“…[30]. The reproducibility and the robustness of this culture process for MSCs was also demonstrated by Nold et al [75].…”

“…Manual Ficoll gradient centrifugation can now also be replaced by completely automated procedures. An automated cell separation system has been developed and it has been demonstrated to yield significantly enhanced cell recoveries when it was compared to the manual Ficoll separation [29,30]. Nevertheless, to facilitate GMP compliance, it is mandatory to reduce at the minimum all the manipulation necessary to produce any cellular product.…”

Engineering mesenchymal stem cells to improve their exosome efficacy and yield for cell‐free therapy

“…Large volumes of media would be required to get a sizable number of vesicles from the MSC-CM that would meet clinical standards. Both the microcarriers and hollow-fibre bioreactors have larger surface area and are currently used for large-scale expansion of MSCs in a 3D environment [51–57] (Figure 4).

10.1080/20013078.2018.1522236-F0004Figure 4.The schematics of spinner flask containing microcarriers containing the cells (A) as well as the hollow-fibre bioreactor and its cross section with cells inside (B).

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Intravenous mesenchymal stromal cell therapy for inflammatory bowel disease: Lessons from the acute graft versus host disease experience

“…In particular, it will be important to identify the types of IBD that may respond to BMSCs, the optimal dose and timing of BMSC infusions and the basis of their immune suppressive activity [118,119]. In addition, whether BMSC treatment will have an unfavorable impact on the anti-pathogen effect and immune responses to infectious agents is a critical issue that needs to be rigorously addressed.…”