Serum-Free Medium for Recombinant Protein Expression in Chinese Hamster Ovary Cells

Liu, Weifeng; Fan, Zhenlin; Lin, Yan; Wang, Tianyun

doi:10.3389/fbioe.2021.646363

Cited by 31 publications

(25 citation statements)

References 93 publications

(110 reference statements)

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“… 1 , 3 , 4 Thus, either complete avoidance of FBS or at least awareness of the effects that some components of FBS might have on experimental outcomes should be considered. 2 , 5 , 6 …”

Section: Introductionmentioning

confidence: 99%

Alkaline Phosphatase Activity of Serum Affects Osteogenic Differentiation Cultures

2022

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Fetal bovine serum (FBS) is a widely used supplement in cell culture medium, despite its known variability in composition, which greatly affects cellular function and consequently the outcome of studies. In bone tissue engineering, the deposited mineralized matrix is one of the main outcome parameters, but using different brands of FBS can result in large variations. Alkaline phosphatase (ALP) is present in FBS. Not only is ALP used to judge the osteogenic differentiation of bone cells, it may affect deposition of mineralized matrix. The present study focused on the enzymatic activity of ALP in FBS of different suppliers and its contribution to mineralization in osteogenic differentiation cultures. It was hypothesized that culturing cells in a medium with high intrinsic ALP activity of FBS will lead to higher mineral deposition compared to media with lower ALP activity. The used FBS types were shown to have significant differences in enzymatic ALP activity. Our results indicate that the ALP activity of the medium not only affected the deposited mineralized matrix but also the osteogenic differentiation of cells as measured by a changed cellular ALP activity of human-bone-marrow-derived mesenchymal stromal cells (hBMSCs). In media with low inherent ALP activity, the cellular ALP activity was increased and played the major role in the mineralization process, while in media with high intrinsic ALP activity contribution from the serum, less cellular ALP activity was measured, and the ALP activity of the medium also contributed to mineral formation substantially. Our results highlight the diverse effects of ALP activity intrinsic to FBS on osteogenic differentiation and matrix mineralization and how FBS can determine the experimental outcomes, in particular for studies investigating matrix mineralization. Once again, the need to replace FBS with more controlled and known additives is highlighted.

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“… 1 , 3 , 4 Thus, either complete avoidance of FBS or at least awareness of the effects that some components of FBS might have on experimental outcomes should be considered. 2 , 5 , 6 …”

Section: Introductionmentioning

confidence: 99%

Alkaline Phosphatase Activity of Serum Affects Osteogenic Differentiation Cultures

2022

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“…Culture medium is another key factor for cell cultures and vaccine yield and quality (Li et al 2021 ). In industrial applications, the optimal medium for biological products must support high cell density, high cell productivity, and reduce the manufacturing costs (Kim et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

et al. 2022

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Herpes zoster is caused by reactivation of the varicella zoster virus (VZV). Researching and developing a herpes zoster vaccine will help to decrease the incidence of herpes zoster. To increase the bioreactor productivity, a serum-free HEK293 cell perfusion process with adenovirus vector herpes zoster (rAd-HZ) vaccine production was developed efficiently using the design of experiment (DoE) method. First, serum-free media for HEK293 cells were screened in both batch and semi-perfusion culture modes. Then, three optimal media were employed in a medium mixture design to improve cell culture performance, and the 1:1 mixture of HEK293 medium and MCD293 medium (named HM293 medium) was identified as the optimal formulation. On the basis of the HM293 medium, the relationship of critical process parameters (CPPs), including the time of infection (TOI), multiplicity of infection (MOI), pH, and critical quality attributes (CQAs) (adenovirus titer (Titer), cell-specific virus yield (CSVY), adenovirus fold expansion (Fold)) of rAd-HZ production was investigated using the DoE approach. Furthermore, the robust setpoint and design space of these CPPs were explored. Finally, the rAd-HZ production process with parameters at a robust setpoint (TOI = 7.2 × 106 cells/mL, MOI = 3.7, and pH = 7.17) was successfully scaled-up to a 3-L bioreactor with an alternating tangential flow system, yielding an adenovirus titer of 3.0 × 1010 IFU/mL, a CSVY of 4167 IFU/cells, a Fold of 1117 at 2 days post infection (dpi). The DoE approach accelerated the development of a HEK293 serum-free medium and of a robust adenovirus production process.

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“…However, the use of FBS in in vitro cell culture is controversial due to a number of reasons, including ethical concerns, shortage in global supply and most importantly its undefined, complex composition and variability which could lead to unexpected and/or unreliable experimental outcomes [1, 3,4]. Thus, either complete avoidance of FBS or at least awareness of the effects that some components of FBS might have on experimental outcomes should be considered [2,5,6]. FBS has previously been described having various effects on mineral deposition.…”

Section: Introductionmentioning

confidence: 99%

Alkaline phosphatase activity of serum affects osteogenic differentiation cultures

Ansari

Ito

Hofmann

2021

Preprint

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Fetal bovine serum (FBS) is a widely used supplement in cell culture medium, despite its known variability in composition which greatly affects cellular function and consequently the outcome of studies. In bone tissue engineering, the deposited mineralized matrix is one of the main outcome parameters, but using different brands of FBS can result in large variations. Alkaline phosphatase (ALP) is present in FBS. Not only is ALP used to judge the osteogenic differentiation of bone cells, it may affect deposition of mineralized matrix. The present study focused on the enzymatic activity of ALP in FBS of different suppliers and its contribution to mineralization in osteogenic differentiation cultures. It was hypothesized that culturing cells in a medium with high intrinsic ALP activity of FBS will lead to higher mineral deposition compared to media with lower ALP activity. The used FBS types were shown to have significant differences in enzymatic ALP activity. Our results indicate that the ALP activity of the medium not only affected the deposited mineralized matrix but also the osteogenic differentiation of cells as measured by a changed cellular ALP activity of human bone marrow derived mesenchymal stromal cells (hBMSC). In media with low inherent ALP activity, the cellular ALP activity was increased and played the major role in the mineralization process; while, in media with high intrinsic ALP activity contribution from the serum, less cellular ALP activity was measured and the ALP activity of the medium also contributed to mineral formation substantially. Our results highlight the diverse effects of ALP activity intrinsic to FBS on osteogenic differentiation and matrix mineralization and how FBS can determine the experimental outcomes, in particular for studies investigating matrix mineralization. Once again, the need to replace FBS with more controlled and known additives is highlighted.

show abstract

Serum-Free Medium for Recombinant Protein Expression in Chinese Hamster Ovary Cells

Cited by 31 publications

References 93 publications

Alkaline Phosphatase Activity of Serum Affects Osteogenic Differentiation Cultures

Alkaline Phosphatase Activity of Serum Affects Osteogenic Differentiation Cultures

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

Alkaline phosphatase activity of serum affects osteogenic differentiation cultures

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