Effects of microcarriers addition and mixing on WJ-MSC culture in bioreactors

Sion, Caroline; Loubière, Céline; Włodarczyk‐Biegun, Małgorzata K.; Davoudi, Neda; Müller‐Renno, Christine; Guédon, Emmanuel; Chevalot, Isabelle; Olmos, Éric

doi:10.1016/j.bej.2020.107521

Cited by 13 publications

(20 citation statements)

References 37 publications

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“…However, despite the percentage of cell colonization on microcarriers being satisfactory, some empty microcarriers were still visible during the culture (Figure 2). It was also noticed that microcarrier aggregates appeared on Day 7, and increased in size, as observed on Day 11 even if it could be assumed that these aggregates were smaller in size in comparison with the no microcarrier feed process (Sion et al, 2020). However, as shown on the live/dead image on Day 11 (Figure 2), some microcarriers were occupied by dead cells and, as demonstrated in a previous study on Cytodex‐1, they were no longer usable by cells (Sion et al, 2020).…”

Section: Resultsmentioning

confidence: 99%

“…It was also noticed that microcarrier aggregates appeared on Day 7, and increased in size, as observed on Day 11 even if it could be assumed that these aggregates were smaller in size in comparison with the no microcarrier feed process (Sion et al, 2020). However, as shown on the live/dead image on Day 11 (Figure 2), some microcarriers were occupied by dead cells and, as demonstrated in a previous study on Cytodex‐1, they were no longer usable by cells (Sion et al, 2020). Thus, considering that these particles became inert could promote damaging collisions on the other active microcarriers, it could be interesting to remove these empty microcarriers using an adapted perfusion system.…”

Section: Resultsmentioning

confidence: 99%

“…Cell extraction and primary cell culture of WJ‐MSCs were performed as described in a previously published work (Sion et al, 2020). Corning Synthemax II‐coated dissolvable microcarriers (DM‐SII) (Corning Cat No.…”

Section: Methodsmentioning

confidence: 99%

“…The maximal number of cells was obtained between Days 9 and 10, with a mean maximal cell concentration of cells of 700 million approximately. In a perfusedcontinuous mode with selective microcarrier removal, the expansion factor was 60 ± 17, whereas it was only of 43 ± 3 in fed-batch mode, and 1.5 in spinner vessels (Sion et al, 2020). In comparison, dos These markers were chosen based on literature results with a demonstrated role in migration, immune modulation, or proliferation properties.…”

Section: Characterization Of Cell Growthmentioning

confidence: 99%

“…Feeding strategies were proposed to provide nutrients supply and to limit concentration of metabolic products to non‐inhibitory levels. In this study, the fed‐batch mode used by Lawson et al (2017) was implemented with the microcarrier feed strategy developed in a previous study (Sion et al, 2020). Finally, “classic” perfusion modes were also tested for the expansion of MSCs.…”

Section: Introductionmentioning

confidence: 99%

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A new perfusion mode of culture for WJ‐MSCs expansion in a stirred and online monitored bioreactor

Sion

Ghannoum

Ebel

et al. 2021

Biotech & Bioengineering

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As a clinical dose requires a minimum of 10 6 cells per kilogram of patients, it is, therefore, crucial to develop a scalable method of production of Wharton Jelly mesenchymal stem cells (WJ-MSCs) with maintained inner characteristics. Scalable expansion of WJ-MSCs on microcarriers usually found in cell culture, involves specific cell detachment using trypsin and could have harmful effects on cells. In this study, the performance of batch, fedbatch, and perfused-continuous mode of culture were compared. The batch and fedbatch modes resulted in expansion factors of 5 and 43, respectively. The perfusedcontinuous mode strategy consisted of the implementation of a settling tube inside the bioreactor. The diameter of the tube was calculated to maintain microcarriers colonized by cells in the bioreactor whereas empty microcarriers (responsible for potentially damaging collisions) were removed, using a continuous flow rate based on MSCs physiological requirements. Thanks to this strategy, a maximal number of 800 million cells was obtained in a 1.5 L bioreactor in 10 days. Lastly, online dielectric spectroscopy was implemented in the bioreactor and indicated that cell growth could be monitored during the culture.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Characterization Of Cell Growthmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A new perfusion mode of culture for WJ‐MSCs expansion in a stirred and online monitored bioreactor

Sion

Ghannoum

Ebel

et al. 2021

Biotech & Bioengineering

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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

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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.

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Impact of microcarrier concentration on mesenchymal stem cell growth and death: Experiments and modeling

Maillot

Isla

Loubière

et al. 2022

Biotech & Bioengineering

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Mesenchymal stem cell (MSC) products are promising therapeutic candidates to treat a wide range of pathologies. The successful commercialization of these cell therapies will, however, depend on the development of reproducible cell production processes. For this, using microcarriers as growth supports within controlled conditions may be a viable process option. Although increasing microcarrier concentration may be associated with greater productivity due to the increased available culture surface, additional friction or shocks between microcarriers are likely to lead to undesired cell death. However, data detailing the impact of microcarrier collisions on MSC growth remains scarce. The following work demonstrates that MSC growth on microcarriers is greatly influenced by particle concentration even when little impact is observed on the apparent growth rate. It is suggested that the apparent growth rate may result in an equilibrium between growth and death kinetics which are independently affected by particle concentration and that certain MSC quality attributes may be progressively degraded in parallel. In addition, the theoretical reduction of the MSC growth rate was modeled according to the ratio between the average interparticle distance and the Kolmogorov scale. This study is an original contribution toward understanding the hydrodynamic effects in microcarrier‐based stem cell cultures.

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Effects of microcarriers addition and mixing on WJ-MSC culture in bioreactors

Cited by 13 publications

References 37 publications

A new perfusion mode of culture for WJ‐MSCs expansion in a stirred and online monitored bioreactor

A new perfusion mode of culture for WJ‐MSCs expansion in a stirred and online monitored bioreactor

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

Impact of microcarrier concentration on mesenchymal stem cell growth and death: Experiments and modeling

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