Agitation in a microcarrier-based spinner flask bioreactor modulates homeostasis of human mesenchymal stem cells

Jeske, Richard; Lewis, S.A.; Tsai, Ang‐Chen; Sanders, Kevin E.; Liu, Chang; Yuan, Xuegang; Li, Yan

doi:10.1016/j.bej.2021.107947

Cited by 14 publications

(5 citation statements)

References 56 publications

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“…At this time, the rotation speed was the minimum to suspend the microcarriers. The rotation speed can be increased without intensifying the shear force and affecting cell proliferation on the microcarriers (Jeske et al 2021 ) to achieve the weightless suspension of the microcarriers, reduce the speed as much as possible (LIOVIC et al 2012 ), and ensure the normal proliferation of cells while avoiding shedding.…”

Section: Resultsmentioning

confidence: 99%

Physical optimization of cell proliferation and differentiation using spinner flask and microcarriers

Yang

Wang

et al. 2022

AMB Expr

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The traditional breeding industry has been increasingly saturated and caused environmental pollution, disease transmission, excessive resource use, and methane emission; however, it still cannot meet the needs of the growing population. To explore other alternatives, researchers focused on cell agriculture and cell-based meat, especially large-scale cell culture. As a prerequisite for production, large-scale culture technology has become an important bottleneck restricting cell-based meat industrialization. In this study, the single-factor variable method was adopted to examine the influence of Cytodex1 microcarrier pretreatment, spinner flask reaction vessel, cell culture medium, serum and cell incubation, and other influencing factors on large-scale cell cultures to identify the optimization parameters suitable for 3D culture environment. Collagen and 3D culture were also prospectively explored to promote myogenesis and cultivate tissue-like muscle fibers that contract spontaneously. This research lays a theoretical foundation and an exploratory practice for large-scale cell cultures and provides a study reference for the microenvironment of myoblast culture in vitro, a feasible direction for the cell therapy of muscular dystrophy, and prerequisites for the industrialized manufacturing of cell-based meat. Graphical Abstract

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Section: Resultsmentioning

confidence: 99%

Physical optimization of cell proliferation and differentiation using spinner flask and microcarriers

Yang

Wang

et al. 2022

AMB Expr

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“…In particular, microcarrier cultures had higher expression of canonical glycolysis related‐genes and lower autophagy‐related genes than the aggregate cultures. VWBRs have 10‐fold lower shear stress than stirred tank bioreactors or spinner flasks (Borys et al., 2021 ; Jeske et al., 2021 , 2023 ), which may be beneficial for EV biogenesis in microcarrier cultures. In addition, the aggregation‐induced integrated stress response (Bijonowski et al., 2020 ) and the possible nutrient diffusion limitation in the aggregate culture may expose the hiPSCs to more cellular stress than microcarrier culture.…”

Section: Discussionmentioning

confidence: 99%

“…Vertical‐Wheel bioreactors (VWBRs) have previously been used for the production of hMSCs on microcarriers (Lembong et al., 2020 ), hPSC aggregates, hPSCs on microcarriers (Borys et al., 2021 ; Cuesta‐Gomez et al., 2023 ; Nogueira et al., 2021 ; Rodrigues et al., 2018 ; Vallabhaneni et al., 2023 ) and cerebellar spheroids (Silva et al., 2020 , 2021 ). The unique design of the hydrodynamics in VWBRs results in 10‐fold lower shear stress levels compared to traditional stirred tank bioreactors (0.1–0.3 dyn/cm 2 vs. 1–3 dyn/cm 2 , respectively) (Borys et al., 2021 , 2020 ; Jeske et al., 2021 , 2023 ). However, the use of VWBRs and the influence of different 3‐D hPSC organization for hPSC‐EV production has not been investigated to date.…”

Section: Introductionmentioning

confidence: 99%

Extracellular vesicle biogenesis of three‐dimensional human pluripotent stem cells in a novel Vertical‐Wheel bioreactor

Muok,

Sun,

Esmonde

et al. 2024

J of Extracellular Bio

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Extracellular vesicles (EVs) secreted by human‐induced pluripotent stem cells (hiPSCs) have great potential as cell‐free therapies in various diseases, including prevention of blood–brain barrier senescence and stroke. However, there are still challenges in pre‐clinical and clinical use of hiPSC‐EVs due to the need for large‐scale production of a large quantity. Vertical‐Wheel bioreactors (VWBRs) have design features that allow the biomanufacturing of hiPSC‐EVs using a scalable aggregate or microcarrier‐based culture system under low shear stress. EV secretion by undifferentiated hiPSCs expanded as 3‐D aggregates and on Synthemax II microcarriers in VWBRs were investigated. Additionally, two types of EV collection media, mTeSR and HBM, were compared. The hiPSCs were characterized by metabolite and transcriptome analysis as well as EV biogenesis markers. Protein and microRNA cargo were analysed by proteomics and microRNA‐seq, respectively. The in vitro functional assays of microglia stimulation and proliferation were conducted. HiPSCs expanded as 3‐D aggregates and on microcarriers had comparable cell number, while microcarrier culture had higher glucose consumption, higher glycolysis and lower autophagy gene expression based on mRNA‐seq. The microcarrier cultures had at least 17–23 fold higher EV secretion, and EV collection in mTeSR had 2.7–3.7 fold higher yield than HBM medium. Microcarrier culture with mTeSR EV collection had a smaller EV size than other groups, and the cargo was enriched with proteins (proteomics) and miRNAs (microRNA‐seq) reducing apoptosis and promoting cell proliferation (e.g. Wnt‐related pathways). hiPSC‐EVs demonstrated the ability of stimulating proliferation and M2 polarization of microglia in vitro. HiPSC expansion on microcarriers produces much higher yields of EVs than hiPSC aggregates in VWBRs. EV collection in mTeSR increases yield compared to HBM. The biomanufactured EVs from microcarrier culture in mTeSR have exosomal characteristics and are functional in microglia stimulation, which paves the ways for future in vivo anti‐aging study.

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“…Stem cells expanded by microcarriers show advantages not only in the large-scale with a relatively low cost but also in the quality and reliability of the stem cells, including less heterogeneity [ 116 ] and better function in the downstream applications [ 117 , 118 , 119 ], compared to the ones expanded by the 2D cell culture systems. In addition, differences in morphology, surface markers, gene expression profiles, and secretome are observed between the microcarrier and conventional 2D systems in the stem cell culture [ 116 , 120 ]. The other characteristic of microcarrier cell culture is that cells can go through a bead-to-bead transfer [ 121 ], which simplifies the subculture process in the large-scale production of stem cells.…”

Section: Current 3d and Dynamic Cell Culture Approachesmentioning

confidence: 99%

Current Advances in 3D Dynamic Cell Culture Systems

Huang

Gao

et al. 2022

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The traditional two-dimensional (2D) cell culture methods have a long history of mimicking in vivo cell growth. However, these methods cannot fully represent physiological conditions, which lack two major indexes of the in vivo environment; one is a three-dimensional 3D cell environment, and the other is mechanical stimulation; therefore, they are incapable of replicating the essential cellular communications between cell to cell, cell to the extracellular matrix, and cellular responses to dynamic mechanical stimulation in a physiological condition of body movement and blood flow. To solve these problems and challenges, 3D cell carriers have been gradually developed to provide a 3D matrix-like structure for cell attachment, proliferation, differentiation, and communication in static and dynamic culture conditions. 3D cell carriers in dynamic culture systems could primarily provide different mechanical stimulations which further mimic the real in vivo microenvironment. In this review, the current advances in 3D dynamic cell culture approaches have been introduced, with their advantages and disadvantages being discussed in comparison to traditional 2D cell culture in static conditions.

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Agitation in a microcarrier-based spinner flask bioreactor modulates homeostasis of human mesenchymal stem cells

Cited by 14 publications

References 56 publications

Physical optimization of cell proliferation and differentiation using spinner flask and microcarriers

Physical optimization of cell proliferation and differentiation using spinner flask and microcarriers

Extracellular vesicle biogenesis of three‐dimensional human pluripotent stem cells in a novel Vertical‐Wheel bioreactor

Current Advances in 3D Dynamic Cell Culture Systems

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