Large-Scale Clinical Expansion of Mesenchymal Stem Cells in the GMP-Compliant, Closed Automated Quantum® Cell Expansion System: Comparison with Expansion in Traditional T-Flasks

Lechanteur, Chantal; Baila, Stefano; Janssens, Michiel Etienne; Giet, Olivier; Briquet, Alexandra; Baudoux, Étienne; Béguin, Yves

doi:10.4172/2157-7633.1000222

Cited by 42 publications

(30 citation statements)

References 31 publications

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“…Immunophenotyping showed that hASCs expanded in flask or HFBR retained a characteristic MSC immunophenotypic profile. In this sense hASCs from both cell expansion systems were remarkably similar, confirming data from the literature obtained with bone marrow-derived MSCs expanded in the same HFBR used in this study [43,44]. Concomitantly, the contamination of endothelial and hematopoietic cells was retained low enough to consider the cell therapy product for both cell expansion systems as pure [18].…”

Section: Discussionsupporting

confidence: 77%

Chondrogenic potential of hASCs expanded in flask or in a hollow-fiber bioreactor

Pirrone¹,

Gobbetti²,

Caprara³

et al. 2017

J Stem Cell Res Med

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In recent years, various clinical trials are exploring the efficacy of cell-based strategies to treat cartilage defects. Human adipose-derived stem/stromal cells (hASCs) have great potential for the regeneration of articular cartilage due to their ability to undergo differentiation into the chondrogenic lineage. In most cell-based therapies, the single dose requires 10 7 -10 9 cells, and then it is necessary to move from manual flask-based culture to bioreactor whose monitoring and control of biological variability is necessary. Furthermore, before clinical use, the success of hASC differentiation need to be thoroughly inspected. In this study, we sought to assess with complementary methods the chondrogenic potential of hASCs expanded in flask or in a hollow-fiber bioreactor (HFBR). Human ASCs, after phenotypic characterization were subjected to chondrogenic differentiation, the expression of cartilage-specific genes was analysed by quantitative real-time polymerase chain reaction (qPCR); optical microscopy as well as transmission electron microscopy (TEM) were also performed. The two culture expansion systems do not affect hASC immunophenotype; microscopy analysis confirmed the successful of hASC differentiation process. Gene expression analysis after differentiation displayed an upregulation of cartilage-specific genes more noticeable in hASCs expanded in the HFBR, suggesting that hASCs expanded in the HFBR can be efficiently differentiated into chondroblasts. Our study demonstrates that flask and HFBR expansion methods do not affect the subsequent differentiation process; additionally, our results highlight the importance of analyzing differentiation by different techniques and support the importance of TEM analysis that is able to give additional critical information to molecular biology results.

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Section: Discussionsupporting

confidence: 77%

Chondrogenic potential of hASCs expanded in flask or in a hollow-fiber bioreactor

Pirrone¹,

Gobbetti²,

Caprara³

et al. 2017

J Stem Cell Res Med

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“…In the current study, genipin cross‐linked alginate–chitosan microcarriers were developed for MSC expansion. The first objective of this article was to characterize and optimize the microcarrier production process to create a microcarrier stable within cell culture environments for 2 weeks – the typical amount of time for stem cell expansion . The second objective was to investigate the suitability of the created microcarriers for the cultivation of MSCs.…”

Section: Introductionmentioning

confidence: 99%

Electrosprayed genipin cross‐linked alginate–chitosan microcarriers for ex vivo expansion of mesenchymal stem cells

Chui

Odeleye

Linh

et al. 2018

J Biomedical Materials Res

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Mesenchymal stem cells (MSCs) are potential therapeutic candidates, owing to their ability to differentiate into several cell types. However, the gap between availability and demand of MSCs requires alternative expansion methods from 2D culture flasks. Microcarriers are a promising approach for MSC expansion due to their high surface area-to-volume ratio. However, current commercial microcarriers do not provide the highest cell yield due to low cell attachment efficiencies and difficulty in cell detachment. This study developed a hydrogel-based microcarrier from genipin cross-linked alginate-chitosan beads. Alginate beads were produced by electrospraying before being coated with chitosan and cross-linked in genipin. The degree of cross-linking was determined through fluorescence reading of the genipin-chitosan conjugates. MSCs cultured on these microcarriers had a 26% higher cell attachment and twice the proliferation rate compared to the commercial microcarrier Cytodex 1. Cells easily detached from the hydrogel beads and did not require extended incubation periods or intense agitation during cell harvest. There was no significant difference in gene expression between the two microcarriers for the positive MSC surface markers CD-90, CD-105, and CD-73 as well as showing either low or no signal for negative MSC surface markers CD-45 and CD-34.

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“…The PDs achieved in Quantum systems were twice as high as in flasks. Studies have compared culture expansion of BM-MSC in flasks vs Quantum systems, and have reported twice as high a BM-MSC yield after two passages compared to flask cultivation [13, 14]. However, two other studies showed no significant differences in PDs when comparing culture expansion of BM-MSC in flasks and Quantum systems [12, 21].…”

Section: Discussionmentioning

confidence: 99%

“…Several studies that have used Quantum systems for culture expansion of BM-MSCs, have used fibronectin as a coating material [11–14, 21]. However, clinical grade fibronectin can be very costly.…”

Section: Discussionmentioning

confidence: 99%

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Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture

et al. 2016

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BackgroundAdipose derived stromal cells (ASCs) are a rich and convenient source of cells for clinical regenerative therapeutic approaches. However, applications of ASCs often require cell expansion to reach the needed dose. In this study, cultivation of ASCs from stromal vascular fraction (SVF) over two passages in the automated and functionally closed Quantum Cell Expansion System (Quantum system) is compared with traditional manual cultivation.MethodsStromal vascular fraction was isolated from abdominal fat, suspended in α-MEM supplemented with 10% Fetal Bovine Serum and seeded into either T75 flasks or a Quantum system that had been coated with cryoprecipitate. The cultivation of ASCs from SVF was performed in 3 ways: flask to flask; flask to Quantum system; and Quantum system to Quantum system. In all cases, quality controls were conducted for sterility, mycoplasmas, and endotoxins, in addition to the assessment of cell counts, viability, immunophenotype, and differentiation potential.ResultsThe viability of ASCs passage 0 (P0) and P1 was above 96%, regardless of cultivation in flasks or Quantum system. Expression of surface markers and differentiation potential was consistent with ISCT/IFATS standards for the ASC phenotype. Sterility, mycoplasma, and endotoxin tests were consistently negative. An average of 8.0 × 107 SVF cells loaded into a Quantum system yielded 8.96 × 107 ASCs P0, while 4.5 × 106 SVF cells seeded per T75 flask yielded an average of 2.37 × 106 ASCs—less than the number of SVF cells seeded. ASCs P1 expanded in the Quantum system demonstrated a population doubling (PD) around 2.2 regardless of whether P0 was previously cultured in flasks or Quantum, while ASCs P1 in flasks only reached a PD of 1.0. Conclusion: Manufacturing of ASCs in a Quantum system enhances ASC expansion rate and yield significantly relative to manual processing in T-flasks, while maintaining the purity and quality essential to safe and robust cell production. Notably, the use of the Quantum system entails significantly reduced working hours and thereby costs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-1080-9) contains supplementary material, which is available to authorized users.

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Large-Scale Clinical Expansion of Mesenchymal Stem Cells in the GMP-Compliant, Closed Automated Quantum® Cell Expansion System: Comparison with Expansion in Traditional T-Flasks

Cited by 42 publications

References 31 publications

Chondrogenic potential of hASCs expanded in flask or in a hollow-fiber bioreactor

Chondrogenic potential of hASCs expanded in flask or in a hollow-fiber bioreactor

Electrosprayed genipin cross‐linked alginate–chitosan microcarriers for ex vivo expansion of mesenchymal stem cells

Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture

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