Quality by design to define critical process parameters for mesenchymal stem cell expansion

Maillot, Charlotte; Sion, Caroline; Isla, Natalia de; Toye, Dominique; Olmos, Éric

doi:10.1016/j.biotechadv.2021.107765

Cited by 14 publications

(11 citation statements)

References 119 publications

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“…Implementing QbD to biopharmaceuticals is especially challenging due to their intrinsic complexity [ 24 ]. On that note, Maillot and co-workers reported that the completion of clinical trials using human mesenchymal stem cells (hMSCs) as therapeutic products must face many hurdles, possibly due to suboptimal quality standards early during development [ 25 ]. In this context, the wider implementation of QbD is pivotal in reaching industrial production for CTPs [ 21 ].…”

Section: Quality-by-design (Qbd) Principlesmentioning

confidence: 99%

“…In the last decade, several studies focused on applying QbD to cell culture, both as an upstream process for the production of specific proteins (e.g., monoclonal antibodies) [ 26 – 33 ] and, more recently, for CTP manufacturing [ 21 , 25 , 34 – 36 ]. The manufacturing of CTPs for clinical application requires several of the following steps: acquisition or generation of the starting cell type; cultivation; modification; harvest; concentration; purification; formulation, and fill and finish (preparing the CTP at the correct concentration and composition, dispensing it into the final product ‘container’, and any post-fill processing); storage; and shipping of the product [ 21 ].…”

Section: Quality-by-design (Qbd) Principlesmentioning

confidence: 99%

See 1 more Smart Citation

Critical Analysis of cGMP Large-Scale Expansion Process in Bioreactors of Human Induced Pluripotent Stem Cells in the Framework of Quality by Design

et al. 2021

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Section: Quality-by-design (Qbd) Principlesmentioning

confidence: 99%

Section: Quality-by-design (Qbd) Principlesmentioning

confidence: 99%

Critical Analysis of cGMP Large-Scale Expansion Process in Bioreactors of Human Induced Pluripotent Stem Cells in the Framework of Quality by Design

et al. 2021

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“…Owing to the scarcity and heterogeneity of freshly isolated hAMSCs, long-term in vitro expansion is the main method to generate the minimum effective dose [8] . Regretfully, long-term in vitro expansion of MSC will cause inevitable replicative or stress-induced senescence, [9] which is accompanied by an increase in genomic instability and genetic damage accumulation [10] , These changes impair the quality and viability of MSCs [11] , ultimately decreasing the e cacy of MSC therapies. For example, radiation-induced senescence abrogated the protective immunoregulatory effect in a mouse model of sepsis [12] .…”

Section: Introductionmentioning

confidence: 99%

Modulation of mesenchymal stem cells protection against senescence during long-term expansion by the PI3K/Akt signaling

Luo

Zhu

Xiao

2023

Preprint

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Background and Objectives: Therapies using mesenchymal stem cells (MSCs) show immense potential and provide a promising new paradigm for treating previously untreatable diseases. These therapies require direct transplantation of a large number of MSCs obtained by long-term expansion in vitro, resulting in inevitable senescence and a decline in MSC quality, characterized by the appearance of senescence signatures, loss of proliferation, and decreased differentiation potential. However, the regulatory mechanism underlying MSC senescence remains unclear. We investigated this mechanism using young (passage 4 [P4]) and aging (P10) human amniotic MSCs (hAMSCs). Methods and Results: P10 hAMSCs showed a senescence phenotype in vitro, including G1-phase cell cycle arrest and increased β-galactosidase-positive staining compared to P4 hAMSCs. Senescence is accompanied by the degeneration of stemness properties, including decreased expression of stemness transcription factors and decreased ability to differentiate into osteoblasts. Further analysis showed that aging hAMSCs contained lower levels of phosphorylated PI3K and AKT proteins, and thus lower activity levels, than those in young hAMSCs. To clarify whether low PI3K/Akt signaling promotes hAMSC senescence, we treated young hAMSCs with a specific inhibitor of phosphorylated AKT, MK2206. The treated cells showed a senescent phenotype at 72 h, accompanied by G1-phase cell cycle arrest and a decrease in proliferative and osteogenic capacities. Conclusions: These data suggest that the PI3K/Akt signaling pathway protects against senescence during long-term in vitro expansion of hAMSCs and plays a central role in maintaining the hAMSC pluripotency.

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“…During the expansion of MSCs in vitro , there are some uncontrolled factors, such as the limited number of primary MSCs [ 2 , 10 ], the decreases of proliferation and differentiation capacity of MSCs after passaging, and the interdonor variability and scalability associated with primary donor-derived MSCs [ 4 , 11 , 12 ], which may affect the number and quality of MSCs before clinical utility [ 1 , 2 , 13 ]. During the transplantation of MSCs, there are still various unsolved problems, such as the loss of MSCs, low homing and engraftment rates of MSCs, inability of precise regulation of MSC differentiation, and accelerated senescence and apoptosis of MSCs after cell transplantation, which are major issues in MSC preclinical research and may influence the clinical utility of MSCs [ 1 – 3 ].…”

Section: Introductionmentioning

confidence: 99%

Effects of Ginsenoside Rg1 on the Biological Behavior of Human Amnion-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs)

Han

Huang

et al. 2023

Stem Cells International

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Ginsenoside Rg1 (Rg1) is purified from ginseng with various pharmacological effects, which might facilitate the biological behavior of human amnion-derived mesenchymal stem/stromal cells (hAD-MSCs). This study is aimed at investigating the effects of Rg1 on the biological behavior, such as viability, proliferation, apoptosis, senescence, migration, and paracrine, of hAD-MSCs. hAD-MSCs were isolated from human amnions. The effects of Rg1 on the viability, proliferation, apoptosis, senescence, migration, and paracrine of hAD-MSCs were detected by CCK-8, EdU, flow cytometry, SA-β-Gal staining, wound healing, and ELISA assays, respectively. The protein expression levels were detected by western blot. Cell cycle distribution was evaluated using flow cytometry. We found that Rg1 promoted hAD-MSC cycle progression from G0/G1 to S and G2/M phases and significantly increased hAD-MSC proliferation rate. Rg1 activated PI3K/AKT signaling pathway and significantly upregulated the expressions of cyclin D, cyclin E, CDK4, and CDK2 in hAD-MSCs. Inhibition of PI3K/AKT signaling significantly downregulated the expressions of cyclin D, cyclin E, CDK4, and CDK2, prevented cell cycle progression, and reduced hAD-MSC proliferation induced by Rg1. hAD-MSC senescence rate was significantly increased by D-galactose, while the elevated hAD-MSC senescence rate induced by D-galactose was significantly decreased by Rg1 treatment. D-galactose significantly induced the expressions of senescence markers, p16INK4a, p14ARF, p21CIP1, and p53 in hAD-MSCs, while Rg1 significantly reduced the expressions of those markers induced by D-galactose in hAD-MSCs. Rg1 significantly promoted the secretion of IGF-I in hAD-MSCs. Rg1 reduced the hAD-MSC apoptosis rate. However, the difference was not significant. Rg1 had no influence on hAD-MSC migration. Altogether, our results demonstrate that Rg1 can promote the viability, proliferation, and paracrine and relieve the senescence of hAD-MSCs. PI3K/AKT signaling pathway is involved in the promotive effect of Rg1 on hAD-MSC proliferation. The protective effect of Rg1 on hAD-MSC senescence may be achieved via the downregulation of p16INK4A and p53/p21CIP1 pathway.

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Quality by design to define critical process parameters for mesenchymal stem cell expansion

Cited by 14 publications

References 119 publications

Critical Analysis of cGMP Large-Scale Expansion Process in Bioreactors of Human Induced Pluripotent Stem Cells in the Framework of Quality by Design

Critical Analysis of cGMP Large-Scale Expansion Process in Bioreactors of Human Induced Pluripotent Stem Cells in the Framework of Quality by Design

Modulation of mesenchymal stem cells protection against senescence during long-term expansion by the PI3K/Akt signaling

Effects of Ginsenoside Rg1 on the Biological Behavior of Human Amnion-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs)

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