Large‐scale culture of a megakaryocytic progenitor cell line with a single‐use bioreactor system

Nurhayati, Retno Wahyu; Ojima, Yoshihiro; Dohda, Takeaki; Kino‐oka, Masahiro

doi:10.1002/btpr.2595

Cited by 10 publications

(8 citation statements)

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“…Influence of shear stress on cell damage has been investigated not only in bioreactors, but also in microfluid devices and blood vessels (Mitchell, 2013;Odeleye, 2014;Model, 2014). Although many researchers have tried to clarify how shear stress affects gene expression and physiological conditions (Nurhayati, 2018;Jayagopal, 2019), the kind of cellular systems that sense the effect of shear stress on gene expression still remains unknown. Even if a type of shear stress induced gene expression in one cell type, it may not affect the same in other cell lines (Akimoto, 2000;Novak, 2019).…”

Section: Discussionmentioning

confidence: 99%

Improved cultivation of CHO cells in bioreactor with reciprocal mixing

Ueki

Tansho

Sato

et al. 2020

Preprint

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We have constructed a new bioreactor with reciprocal mixing that is better suited for the cultivation of delicate animal cells. In-silico simulation (computational fluid dynamics) suggested both maximum and average shear stresses in the bioreactor with reciprocal mixing to be remarkably lower than in conventional bioreactor with rotary mixing. Although we could not find any difference in growth speed and cell density between the bioreactors with reciprocal and rotary mixing, we did find cell viability in reciprocal-mixing bioreactor to be retained longer than in rotary-paddle bioreactor. This implied that cell culture in a bioreactor with reciprocal mixing could be prolonged for the production of target proteins. Leakage of lactate dehydrogenase activity into the culture medium was suppressed much more in the reciprocal-mixing bioreactor than in the rotary-paddle one. Production of human tissue plasminogen activator in the former was also observed to be much more than in the latter. Therefore, bioreactor with reciprocal mixing was concluded to be better suited for the cultivation of animal cells and efficient production of proteins, such as antibody drugs and various growth factors.

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Section: Discussionmentioning

confidence: 99%

Improved cultivation of CHO cells in bioreactor with reciprocal mixing

Ueki

Tansho

Sato

et al. 2020

Preprint

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“…We were able to expand imMKCLs effectively in static or rocking-motion type bioreactors [73]. Another group used a single-use bioreactor to expand commercial megakaryocytes through automated gas exchange and orbital shaking, which might be also useful [74].…”

Section: Establishment Of Expandable Megakaryocytic Cell Lines Througmentioning

confidence: 99%

Generation and manipulation of human iPSC-derived platelets

Sugimoto

Eto

2021

Cell. Mol. Life Sci.

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The discovery of iPSCs has led to the ex vivo production of differentiated cells for regenerative medicine. In the case of transfusion products, the derivation of platelets from iPSCs is expected to complement our current blood-donor supplied transfusion system through donor-independent production with complete pathogen-free assurance. This derivation can also overcome alloimmune platelet transfusion refractoriness by resulting in autologous, HLA-homologous or HLA-deficient products. Several developments were necessary to produce a massive number of platelets required for a single transfusion. First, expandable megakaryocytes were established from iPSCs through transgene expression. Second, a turbulent-type bioreactor with improved platelet yield and quality was developed. Third, novel drugs that enabled efficient feeder cell-free conditions were developed. Fourth, the platelet-containing suspension was purified and resuspended in an appropriate buffer. Finally, the platelet product needed to be assured for competency and safety including non-tumorigenicity through in vitro and in vivo preclinical tests. Based on these advancements, a clinical trial has started. The generation of human iPSC-derived platelets could evolve transfusion medicine to the next stage and assure a ubiquitous, safe supply of platelet products. Further, considering the feasibility of gene manipulations in iPSCs, other platelet products may bring forth novel therapeutic measures.

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“…Lactate is commonly accumulated under high cell density and limited oxygen transfer (Ouyang et al, 2007). The stirring factors cause cells to experience hydrodynamic stress (Bergemann et al, 2015;Nurhayati et al, 2018). As illustrated in Figure 6, the 2D culture using a multi-well plate on day 2 revealed that the cells were around 30% confluent.…”

Section: Cell Proliferation In Static 2d and Dynamic 3d Culturesmentioning

confidence: 99%

The Effects of Static and Dynamic Culture Systems on Cell Proliferation and Conditioned Media of Umbilical Cord-derived Mesenchymal Stem Cells

Nurhayati¹,

Lubis²,

Pratama³

et al. 2021

IJTech

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Preclinical and clinical studies have demonstrated the therapeutic effects of umbilical cord-derived mesenchymal stem cells (UC-MSCs) and secretome to cure various degenerative diseases. Thus, the mass-scale production of MSCs is necessary to ensure their availability and costeffectiveness. In the current study, we evaluated the effect of dynamic 3D and static 2D culture systems on cell proliferation and conditioned media of UC-MSCs. The lysate of concentrated thrombocyte was used to substitute animal-derived serum in the culture media. From two experimental sets with different UC and lysates of concentrated thrombocyte donors, it was found that the shortest PDTs for experimental set 1 were 12.3 h (2D culture) and 14.8 h (3D culture), whereas in experimental set 2, they were 17.7 h (2D culture) and 16.9 h (3D culture). Microscopic observation confirmed the formation of cell aggregates in the 3D system, particularly during the exponential phase. SDS-PAGE analysis revealed similar protein profiles of conditioned media from both culture systems. An anti-inflammatory cytokine, namely tumor necrosis factor beta (TGF-β), was analyzed using ELISA to evaluate the effect of culturing methods on TGF-β release. Interestingly, the relative TGF-β contents in the 2D culture were stagnant throughout the incubation times, whereas a higher accumulation of TGF-β was detected in the 3D culture, which was most likely caused by shear stress. Our study confirmed that a dynamic culture system with a microcarriersupported bioreactor is a promising approach to scaling up MSC and secretome productions.

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Large‐scale culture of a megakaryocytic progenitor cell line with a single‐use bioreactor system

Cited by 10 publications

References 50 publications

Improved cultivation of CHO cells in bioreactor with reciprocal mixing

Improved cultivation of CHO cells in bioreactor with reciprocal mixing

Generation and manipulation of human iPSC-derived platelets

The Effects of Static and Dynamic Culture Systems on Cell Proliferation and Conditioned Media of Umbilical Cord-derived Mesenchymal Stem Cells

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