A comprehensive review of quantum bioreactor cell manufacture: Research and clinical applications

Hulme, Charlotte; Mennan, Claire; McCarthy, Helen; Davies, R. L.; Lan, Tian; Rix, Larissa; Perry, Jade; Wright, Karina

doi:10.1016/j.jcyt.2023.04.004

Cited by 6 publications

(4 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this report, we successfully establish a developmental workflow for producing large quantities of EVs from MSCs using a hollow-fiber bioreactor-based system. Importantly this system is reported to be scalable, reproducible, and GMP-compliant for other applications [ 42 , 43 ]. The key advantages for EV production is that it requires relatively small volumes of media for MSC growth, enabling higher yields of EVs per mL and multiple production cycles can be more performed without subculturing compared to conventional flask formats.…”

Section: Discussionmentioning

confidence: 99%

“…tangential flow filtration (TFF), they typically add costs, take more time, and through sheer forces can lead to aggregation and loss of EV integrity (unpublished results). Using UC for downstream purification, there are now GMP-compliant continuous flow UC systems capable of handling 40–100 L of media making this approach more compatible for large volumes needed for clinical trials [ 42 , 43 ]. While ultimately the manufacturing configuration required for mass production depends on the clinical trial, combining a scaled-up version of the bioreactor coupled with newer UC systems may be a cost-effective approach of manufacturing functionally active EVs.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome

Kink,

Bellio,

Forsberg

et al. 2024

Stem Cell Res Ther

View full text Add to dashboard Cite

Background Hematopoietic acute radiation syndrome (H-ARS) occurring after exposure to ionizing radiation damages bone marrow causing cytopenias, increasing susceptibility to infections and death. We and others have shown that cellular therapies like human mesenchymal stromal cells (MSCs), or monocytes/macrophages educated ex-vivo with extracellular vesicles (EVs) from MSCs were effective in a lethal H-ARS mouse model. However, given the complexity of generating cellular therapies and the potential risks of using allogeneic products, development of an “off-the-shelf” cell-free alternative like EVs may have utility in conditions like H-ARS that require rapid deployment of available therapeutics. The purpose of this study was to determine the feasibility of producing MSC-derived EVs at large scale using a bioreactor and assess critical quality control attributes like identity, sterility, and potency in educating monocytes and promoting survival in a lethal H-ARS mouse model. Methods EVs were isolated by ultracentrifugation from unprimed and lipopolysaccharide (LPS)-primed MSCs grown at large scale using a hollow fiber bioreactor and compared to a small scale system using flasks. The physical identity of EVs included a time course assessment of particle diameter, yield, protein content and surface marker profile by flow-cytometry. Comparison of the RNA cargo in EVs was determined by RNA-seq. Capacity of EVs to generate exosome educated monocytes (EEMos) was determined by qPCR and flow cytometry, and potency was assessed in vivo using a lethal ARS model with NSG mice. Results Physical identity of EVs at both scales were similar but yields by volume were up to 38-fold more using a large-scale bioreactor system. RNA-seq indicated that flask EVs showed upregulated let-7 family and miR-143 micro-RNAs. EEMos educated with LPS-EVs at each scale were similar, showing increased gene expression of IL-6, IDO, FGF-2, IL-7, IL-10, and IL-15 and immunophenotyping consistent with a PD-L1 high, CD16 low, and CD86 low cell surface expression. Treatment with LPS-EVs manufactured at both scales were effective in the ARS model, improving survival and clinical scores through improved hematopoietic recovery. EVs from unprimed MSCs were less effective than LPS-EVs, with flask EVs providing some improved survival while bioreactor EVs provide no survival benefit. Conclusions LPS-EVs as an effective treatment for H-ARS can be produced using a scale-up development manufacturing process, representing an attractive off-the-shelf, cell-free therapy.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome

Kink,

Bellio,

Forsberg

et al. 2024

Stem Cell Res Ther

View full text Add to dashboard Cite

show abstract

“…[9] Alternatives such as cell factories (e.g., HYPERStacks from Corning or Xpansion multiplate bioreactor from Pall) allow an increase in available cell culture surface but may only offer intermediate solutions when compared to 3D systems. Cell culture bioprocessing strategies such as hollow fiber bioreactors (e.g., Quantum Cell Expansion System from Terumo) [10] and microcarriers (MCs), together with vertical wheel bioreactors, [11][12][13] wave bioreactors [14] and stirred-tank (bio)reactors (STRs), provide an increased surface-to-volume ratio in cultures requiring large area attachment surfaces, thus contributing to cost-efficient process designs. Among them, spinner flasks and STRs have been widely investigated, [15][16][17][18][19][20][21][22][23][24] since they are versatile systems and easy to scale up.…”

Section: Introductionmentioning

confidence: 99%

Identification of critical process parameters for expansion of clinical grade human Wharton's jelly‐derived mesenchymal stromal cells in stirred‐tank bioreactors

López‐Fernández,

Garcia‐Gragera,

Lecina

et al. 2024

Biotechnology Journal

View full text Add to dashboard Cite

Cell therapies based on multipotent mesenchymal stromal cells (MSCs) are traditionally produced using 2D culture systems and platelet lysate‐ or serum‐containing media (SCM). Although cost‐effective for single‐dose autologous treatments, this approach is not suitable for larger scale manufacturing (e.g., multiple‐dose autologous or allogeneic therapies with banked MSCs); automated, scalable and Good Manufacturing Practices (GMP)‐compliant platforms are urgently needed. The feasibility of transitioning was evaluated from an established Wharton's jelly MSCs (WJ‐MSCs) 2D production strategy to a new one with stirred‐tank bioreactors (STRs). Experimental conditions included four GMP‐compliant xeno‐ and serum‐free media (XSFM) screened in 2D conditions and two GMP‐grade microcarriers assessed in 0.25 L‐STRs using SCM. From the screening, a XSFM was selected and compared against SCM using the best‐performing microcarrier. It was observed that SCM outperformed the 2D‐selected medium in STRs, reinforcing the importance of 2D‐to‐3D transition studies before translation into clinical production settings. It was also found that attachment efficiency and microcarrier colonization were essential to attain higher fold expansions, and were therefore defined as critical process parameters. Nevertheless, WJ‐MSCs were readily expanded in STRs with both media, preserving critical quality attributes in terms of identity, viability and differentiation potency, and yielding up to 1.47 × 109 cells in a real‐scale 2.4‐L batch.

show abstract

“…We have previously demonstrated that use of the Quantum ® bioreactor (manufactured by Terumo BCT, Lakewood, USA) can generate large numbers of bone marrow- and umbilical cord-derived mesenchymal stromal cells (MSCs), which maintain their phenotypic properties following rapid expansion. 3 This bioreactor system has been widely used to expand human MSCs from various tissue sources, including production of cells used clinically, as we have reviewed and illustrated in Hulme et al 4 We are, however, unaware of any studies that have culture-expanded chondrocytes using this platform. The Quantum ® cell expansion system comprised hollow fibers that provide a surface area of 2.1 m 2 for cell adherence, equivalent to 120 T175 flasks used in standard tissue culture practice.…”

Section: Introductionmentioning

confidence: 99%

The Upscale Manufacture of Chondrocytes for Allogeneic Cartilage Therapies

Hulme

Garcia

Mennan

et al. 2023

Tissue Engineering Part C: Methods

Self Cite

View full text Add to dashboard Cite

Allogeneic chondrocyte therapies need to be developed to allow more individuals to be treated with a cell therapy for cartilage repair and to reduce the burden and cost of the current two-stage autologous procedures. Upscale manufacture of chondrocytes using a bioreactor could help provide an off-the-shelf allogeneic chondrocyte therapy with many doses being produced in a single manufacturing run. In this study, we assess a good manufacturing practice-compliant hollow-fiber bioreactor (Quantum ® ) for adult chondrocyte manufacture. Chondrocytes were isolated from knee arthroplasty-derived cartilage ( n = 5) and expanded in media supplemented with 10% fetal bovine serum (FBS) or 5% human platelet lysate (hPL) on tissue culture plastic (TCP) for a single passage. hPL-supplemented cultures were then expanded in the Quantum bioreactor for a further passage. Matched, parallel cultures in hPL or FBS were maintained on TCP. Chondrocytes from all culture conditions were characterized in terms of growth kinetics, morphology, immunoprofile, chondrogenic potential (chondrocyte pellet assays), and single telomere length analysis. Quantum expansion of chondrocytes resulted in 86.4 ± 38.5 × 10 6 cells in 8.4 ± 1.5 days, following seeding of 10.2 ± 3.6 × 10 6 cells. This related to 3.0 ± 1.0 population doublings in the Quantum bioreactor, compared with 2.1 ± 0.6 and 1.3 ± 1.0 on TCP in hPL- and FBS-supplemented media, respectively. Quantum- and TCP-expanded cultures retained equivalent chondropotency and mesenchymal stromal cell marker immunoprofiles, with only the integrin marker, CD49a, decreasing following Quantum expansion. Quantum-expanded chondrocytes demonstrated equivalent chondrogenic potential (as assessed by ability to form and maintain chondrogenic pellets) with matched hPL TCP populations. hPL manufacture, however, led to reduced chondrogenic potential and increased cell surface positivity of integrins CD49b, CD49c, and CD51/61 compared with FBS cultures. Quantum expansion of chondrocytes did not result in shortened 17p telomere length when compared with matched TCP cultures. This study demonstrates that large numbers of adult chondrocytes can be manufactured in the Quantum hollow-fiber bioreactor. This rapid, upscale expansion does not alter chondrocyte phenotype when compared with matched TCP expansion. Therefore, the Quantum provides an attractive method of manufacturing chondrocytes for clinical use. Media supplementation with hPL for chondrocyte expansion may, however, be unfavorable in terms of retaining chondrogenic capacity. Impact statement This is the first study, to our knowledge, to manufacture adult chondrocytes in a good manufacturing practice-compliant hollow-fiber bioreactor (Quantum ® ). We provide evidence that chondrocytes can be manufactured using this methodology while retaining comparable properties to chondrocytes expanded in matched tissue cul...

show abstract

A comprehensive review of quantum bioreactor cell manufacture: Research and clinical applications

Cited by 6 publications

References 69 publications

Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome

Large-scale bioreactor production of extracellular vesicles from mesenchymal stromal cells for treatment of acute radiation syndrome

Identification of critical process parameters for expansion of clinical grade human Wharton's jelly‐derived mesenchymal stromal cells in stirred‐tank bioreactors

The Upscale Manufacture of Chondrocytes for Allogeneic Cartilage Therapies

Contact Info

Product

Resources

About