Oxygenation in cell culture: Critical parameters for reproducibility are routinely not reported

Al-Ani, Abdullah; Toms, Derek; Kondro, Douglas; Thundathil, Jacob C.; Yu, Yan; Ungrin, Mark

doi:10.1371/journal.pone.0204269

Cited by 114 publications

(117 citation statements)

References 68 publications

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“…However, this was not confirmed with PC-3 cells that were treated with N-(3-mb)-BS, since no significant DNA damage was observed. Most of the data obtained on cell culture, including our own, are performed in 21% oxygen, while the tissue oxygen level is between 3 to 8% and there are some evidences that the high oxygen concentration routinely used in cell culture might lead to altered cellular pathways in vitro [17,18]. Hence, another goal of the current study was to compare the cytotoxicity of N-allyl-BS and N-(3-mb)-BS towards prostate cells that were cultured under 21% and 4% oxygen.…”

Section: Discussionmentioning

confidence: 96%

In Vitro Anti-Prostate Cancer Activity of Two Ebselen Analogues

et al. 2020

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Scientific research has been underway for decades in order to develop an effective anticancer drug, and it has become crucial to find a novel and effective chemotherapeutics in the case of prostate cancer treatment. Ebselen derivatives have been shown to possess a variety of biological activities, including cytostatic and cytotoxic action against tumor cells. In this study, the cytotoxic effect and anticancer mechanism of action of two organoselenium compounds— (N-allyl-1,2-benzisoselenazol-3(2H)-one (N-allyl-BS) and N-(3-methylbutyl)-1,2-benzisoselenazol-3(2H)-one) (N-(3-mb)-BS)—were investigated on two phenotypically different prostate cancer cell lines DU 145 and PC-3. The influence of analyzed compounds on the viability parameter was also assessed on normal prostate cell line PNT1A. The results showed that both organoselenium compounds (OSCs) efficiently inhibited cancer cell proliferation, whereas normal PNT1A cells were less sensitive to the analazyed ebselen analouges. Both OSCs induced G2/M cell cycle arrest and prompted cell death through apoptosis. The detection of cleaved Poly (ADP-ribose) Polymerase (PARP) confirmed this. In addition, N-allyl-BS and N-(3-m)-b-BS increased the level of reactive oxygen species (ROS) formation, however only N-allyl-BS induced DNA damage. Based on our data, we assume that OSCs’ anticancer action can be associated with oxidative stress induction and inactivation of the Akt- dependent signalling pathway. In conclusion, our data demonstrate that ebselen derivatives showed strong cytotoxic efficiency towards prostate cancer cells and may be elucidated as a novel, potent anticancer agent.

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Section: Discussionmentioning

confidence: 96%

In Vitro Anti-Prostate Cancer Activity of Two Ebselen Analogues

et al. 2020

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“…All of these parameters can be manipulated by the frequency of medium changes (Wright Muelas et al, 2018). Medium changes have been shown to influence oxygen supply and cellular metabolism, as well as synthesis of proteins, DNA, and RNA (Al-Ani et al, 2018;Wright Muelas et al, 2018). These observations clearly highlight the necessity for precise documentation and reporting of parameters like cell density and frequency of medium changes to minimize their contribution to insufficient reproducibility.…”

Section: Recommendationsmentioning

confidence: 99%

In Vitro Research Reproducibility: Keeping Up High Standards

2019

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Concern regarding the reproducibility of observations in life science research has emerged in recent years, particularly in view of unfavorable experiences with preclinical in vivo research. The use of cell-based systems has increasingly replaced in vivo research and the application of in vitro models enjoys an ever-growing popularity. To avoid repeating past mistakes, high standards of reproducibility and reliability must be established and maintained in the field of in vitro biomedical research. Detailed guidance documenting the appropriate handling of cells has been authored, but was received with quite disparate perception by different branches in biomedical research. In that regard, we intend to raise awareness of the reproducibility issue among scientists in all branches of contemporary life science research and their individual responsibility in this matter. We have herein compiled a selection of the most susceptible steps of everyday in vitro cell culture routines that have the potential to influence cell quality and recommend practices to minimize the likelihood of poor cell quality impairing reproducibility with modest investment of time and resources.

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“…The correlation between culture medium depth and O 2 delivery to cell cultures has been known for decades. 79 However, it is only recently gaining attention as a contributing factor to poor study reproducibility [80][81][82][83] and presents a particularly important consideration for full vessel culture. Oxygen delivery to cells is governed by two overarching equations: (a) Henry's law of solubility, which describes the solubility of a gas in liquid, and (b) Fick's law of diffusion, which describes the flux of a gas in proportion to the thickness of a fluid layer and concentration gradient across a diffusion barrier.…”

Section: F I G U R Ementioning

confidence: 99%

Factors implicating the validity and interpretation of mechanobiology studies in simulated microgravity environments

Poon

2020

Engineering Reports

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Simulated microgravity (s-μg) devices provide unique conditions for elucidating the effects of gravitational unloading on biological processes and are increasingly being applied for mechanobiology studies. However, without proper characterization of the mechanical environment generated by these systems, the interpretation of results is confounded and limited. Furthermore, the cell culture approaches central to s-μg experimentation introduces new factors that can fundamentally affect results, but these are currently not addressed. It is essential to understand the complete culture environment and how constituent conditions can individually and synergistically affect cellular responses in order to correctly interpret results, otherwise outcomes may be misattributed to the effects of microgravity alone. For the benefit of the growing space biology community, this article critically reviews a typical s-μg cell culture environment in terms of three key conditions: fluid-mediated mechanical stimuli, oxygen tension and biochemical. These and the implications of other experimental variables for biological analysis are categorically discussed. A new set of controls is proposed to properly evaluate the respective effects of these conditions in s-μg culture, along with a reporting matrix and potential strategies for addressing the current limitations of simulated microgravity devices as a platform for mechanobiology research.

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Oxygenation in cell culture: Critical parameters for reproducibility are routinely not reported

Cited by 114 publications

References 68 publications

In Vitro Anti-Prostate Cancer Activity of Two Ebselen Analogues

In Vitro Anti-Prostate Cancer Activity of Two Ebselen Analogues

In Vitro Research Reproducibility: Keeping Up High Standards

Factors implicating the validity and interpretation of mechanobiology studies in simulated microgravity environments

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