Performance evaluation of a proliferation chamber with external stirred conditioning tank for expansion of a suspendable stem cell model

Khosrowshahi, Younes Beygi; Khoshfetrat, Ali Baradar; Abolghasemi, Zahra; Asenjan, Karim Shams

doi:10.1016/j.procbio.2015.04.009

Cited by 6 publications

(4 citation statements)

References 40 publications

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“…Endothelial cells become elongated and alter their morphology, function, and gene expression when they exposure to shear stress [ 35 , 36 ]. We have already shown that the R c parameter may be used to monitor indirectly the stress level exerted on the cells in the dynamic culture systems [ 13 ]. The quantitative morphological analysis of the cells in the present study also revealed that the cell morphology altered more to stretch shape (lower R c values) under the dynamic cultures, particularly at the agitation rate of 100 rpm.…”

Section: Discussionmentioning

confidence: 99%

“…The cell morphology was quantitatively evaluated by a parameter named cell roundness, Rc as described elsewhere [ 13 , 24 ]. Briefly, the images of cells were captured by using a light microscope (Model: BX51; Olympus, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…On the other hand, there is a close relationship between the bioactivity of hematopoietic cells with shear stress, making aeration and agitation rates critical parameters in the STB systems to maintain HSCs functionalities [ 12 ]. To decrease the shear stress in the culture of hematopoietic cells, we have already used the proliferation chamber connected to the STB system as a conditioning vessel [ 3 , 13 ]. Recently, we have also designed a conceptual bioprocess based on the stirred tank bioreactor with a sequencing batch aeration system and evaluated the bioreactor performance for one-step clinical mass production of hematopoietic cells [ 7 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

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Influence of shear force on ex vivo expansion of hematopoietic model cells in a stirred tank bioreactor

et al. 2023

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To evaluate shear stress influence on ex vivo expansion of hematopoietic cell lineages for clinical application, in this study, human pro-monocytic cell (namely U937 cell line) was selected as a hematopoietic stem cell (HSC) model and cultured in suspension mode at two different agitation rates (50, 100 rpm) in the stirred bioreactor. At the agitation rate of 50 rpm, the cells achieved higher expansion folds (27.4 fold) with minimal morphological changes as well as apoptotic cell death, while at 100 rpm the expansion fold decreased after 5-day of culture in suspension culture in comparison with static culture and reached 24.5 fold at the end of the culture. The results of glucose consumption and lactate production were also in agreement with the data of fold expansion and indicated the preference of culture in the stirred bioreactor when agitated at 50 rpm. This study indicated the stirred bioreactor system with an agitation rate of 50 rpm and surface aeration may be used as a potential dynamic culture system for clinical applications of hematopoietic cell lineage. The current experiments shed data related to the effect of shear stress on human U937 cells, as a hematopoietic cell model, to set a protocol for expansion of HSCs for biomedical applications.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Influence of shear force on ex vivo expansion of hematopoietic model cells in a stirred tank bioreactor

et al. 2023

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“…The growth efficiency and homogeneous of cultured SCs in stirring bioreactor were improved in somehow, due to homogeneous nature of culture system. However, SCs are sensitive to shear stress, and agitation as well as aeration affects the cell behaviors such as proliferation and differentiation [11,21].…”

Section: Stirred Tank Bioreactormentioning

confidence: 99%

Application of Bioreactor in Stem Cell Culture

Zhang¹,

Wang²,

Pong³

et al. 2017

JBiSE

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Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applications such as cell therapy, tissue engineering, regenerative medicine and drug screening. However, the clinical applications of SCs require a large amount of SCs with high quality and the number of SCs from their tissue resources is very limited. Large-scale expansion is needed to generate homogeneous SCs with good biological characteristics for clinical application. This necessitates a bioreactor system to provide controllable and stable conditions for stem cell (SC) culture. Traditional methods of bioreactor for maintenance and expansion of cells rely on two-dimensional (2-D) culture techniques, leading to loss self-renewal ability and differentiation capacity upon long-term culture. New approaches for SC expansion with bioreactor employ three-dimensional (3-D) cell growth to mimic their environment in vivo. In this review, we summarize the application of bioreactors in SC culture.

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Ex vivo expansion of hematopoietic stem cells in a proliferation chamber with external stirred conditioning tank: Sequential optimization of growth factors

Khosrowshahi

Khoshfetrat

Shamsasenjan

2015

Engineering in Life Sciences

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The insufficient number of hematopoietic stem cells (HSCs) extracted from various cell sources creates the need for the use of growth factors in the culture systems. Numerous types and concentrations of growth factors have been reported in the literature. In this study, we investigated the effect of three important hematopoietic growth factors thrombopoietin (TPO), stem cell factor (SCF), and Fms‐like tyrosine kinase‐3 ligand (Flt‐3) as well as cell‐seeding density on ex vivo expansion of human HSCs using a factorial design. Sequential optimization was then followed by methods of steepest ascent and central composite design. The optimum concentrations of growth factors were 50, 90, and 34 ng/mL for SCF, TPO, and Flt‐3, respectively, at an initial cell density of 2.5 × 105 cells/mL. Effective expansion factor value (EEF) of HSCs increased considerably and revealed almost similar results when the cells were cultured in a 24‐well plate (EEF = 4.54 ± 0.43) and a proliferation chamber with an external stirred conditioning tank (PC‐ESCT; EEF = 5.1 ± 0.35) at seeding density of 2.5 × 105 cells/mL after 7 days. The cells did not show considerable changes in proliferation when they were cultured in medium containing serum or in a commercial serum‐free medium at the optimum concentrations of the growth factors. The present study demonstrated the optimum condition of hematopoietic growth factors as well as the potential of PC‐ESCT for ex vivo expansion of HSCs.

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Performance evaluation of a proliferation chamber with external stirred conditioning tank for expansion of a suspendable stem cell model

Cited by 6 publications

References 40 publications

Influence of shear force on ex vivo expansion of hematopoietic model cells in a stirred tank bioreactor

Influence of shear force on ex vivo expansion of hematopoietic model cells in a stirred tank bioreactor

Application of Bioreactor in Stem Cell Culture

Ex vivo expansion of hematopoietic stem cells in a proliferation chamber with external stirred conditioning tank: Sequential optimization of growth factors

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