Animal tissue culture principles and applications

Verma, Anju; Verma, Megha; Singh, Akashdeep

doi:10.1016/b978-0-12-811710-1.00012-4

Cited by 98 publications

(76 citation statements)

References 46 publications

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“…Primary cells have several limitations compared to permanent cultures. One is the general need for fresh animal tissue to start laboratory experiments, which may be related to a risk of contamination (Verma et al 2020 ). Moreover, to obtain primary cells, animals have to be sacrificed which is in conflict with ethical considerations.…”

Section: Resultsmentioning

confidence: 99%

In vitro infection of Madin-Darby bovine kidney (MDBK) cells with Eimeria acervulina sporozoites: quantitative analysis of parasite cellular invasion and replication using real-time polymerase chain reaction (PCR)

et al. 2021

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Poultry coccidiosis causes considerable economical losses to the livestock industry. Eimeria parasites are responsible for this disease. On a global scale, E. acervulina and E. tenella are amongst the most common Eimeria spp. infecting broilers. E. tenella is commonly used as infection model in in vivo and in vitro studies. On the other hand, E. acervulina has barely been studied under in vitro conditions. A well established and widely used in vitro model for E. tenella infection is the Madin-Darby bovine kidney cell line (MDBK); however, little is known regarding suitability of MDBK cells as host cells for E. acervulina. We infected MDBK monolayers with two different doses, 5 × 104 and 2 × 105, of E. acervulina sporozoites and evaluated cultures at 24 and 96 h post infection (hpi). For comparison, we ran an identical infection assay using E. tenella sporozoites. To assess parasite reproduction, the number of DNA copies of E. acervulina SCAR marker and E. tenella ITS-1 gene was quantified using real-time quantitative PCR. We found that the number of E. acervulina copies increased significantly at 24 hpi in comparison to E. tenella (p < 0.05). After 96 hpi, E. acervulina gene copies were considerably reduced while E. tenella continued to multiply (p < 0.05). Our results show that MDBK monolayers could be used for in vitro research aimed to study E. acervulina sporozoite cell invasion. Nevertheless, modifications of in vitro cultivation appear necessary to allow qualitative and quantitative studies over longer periods of parasite reproduction.

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Section: Resultsmentioning

confidence: 99%

In vitro infection of Madin-Darby bovine kidney (MDBK) cells with Eimeria acervulina sporozoites: quantitative analysis of parasite cellular invasion and replication using real-time polymerase chain reaction (PCR)

et al. 2021

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“…The FLUX reporter gene can be overexpressed in various types of cell-line such as Human Embryonic Kidney cells (HEK293), Mouse Embryotic Fibroblast cells (NIH3T3), Monkey Kidney Epithelial cells (COS), Human hepatocellular carcinoma cells (HepG2). The first three cell lines are the most commonly used cell lines for biomedical research (Verma et al, 2020). In particular, HEK293 cells and its derivatives have been used extensively in the transfection-based experiments, protein expression, and productions of biologics and pharmaceuticals production (Verma, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…The first three cell lines are the most commonly used cell lines for biomedical research (Verma et al, 2020). In particular, HEK293 cells and its derivatives have been used extensively in the transfection-based experiments, protein expression, and productions of biologics and pharmaceuticals production (Verma, 2014). They have high efficiency of transfection and protein production, and demonstrate reliable translation and processing of protein targets (Thomas & Smart, 2005).…”

Section: Discussionmentioning

confidence: 99%

High Sensitivity and Low-Cost Flavin luciferase (FLUX)-based Reporter Gene for Mammalian Cell Expression

Phonbuppha

Tinikul

Ohmiya

et al. 2021

Preprint

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Luciferase-based gene reporters generating bioluminescence signals are important tools for biomedical research. Amongst the luciferases, flavin-dependent enzymes use the most common, and thus most economical chemicals. However, their applications in mammalian cells are limited due to their low signals compared to other systems. Here, we constructed Flavin Luciferase for Mammalian Cell Expression (FLUX) by engineering luciferase from Vibrio campbellii (the most thermostable bacterial luciferase reported to date) and optimizing its expression and reporter assays in mammalian cells. We found that the FLUX reporter gene can be overexpressed in various cell lines and showed outstanding signal-to-background in HepG2 cells, significantly higher than that of firefly luciferase (Fluc). The combined use of FLUX/Fluc as target/control vectors gave the most stable signals, better than the standard set of Fluc(target)/Rluc(control). We demonstrated that FLUX can be used for testing inhibitors of the NF-kappa-B signaling pathway, validating FLUX applications for various assays in the future.

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“…Carbon dioxide in a cultivated meat bioprocess is closely linked to the cell density, and can readily diffuse across the cell membrane, affecting cellular metabolism and resulting in a lower intracellular pH. The determination of dissolved CO 2 is more difficult due to its chemical reactions with water and the cell culture medium buffered with the CO 2 -bicarbonate based buffer [ 206 ].…”

Section: Sensorsmentioning

confidence: 99%

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production

Djisalov

Knežić

Podunavac

et al. 2021

Biology

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Meat cultivation via cellular agriculture holds great promise as a method for future food production. In theory, it is an ideal way of meat production, humane to the animals and sustainable for the environment, while keeping the same taste and nutritional values as traditional meat and having additional benefits such as controlled fat content and absence of antibiotics and hormones used in the traditional meat industry. However, in practice, there is still a number of challenges, such as those associated with the upscale of cultured meat (CM). CM food safety monitoring is a necessary factor when envisioning both the regulatory compliance and consumer acceptance. To achieve this, a multidisciplinary approach is necessary. This includes extensive development of the sensitive and specific analytical devices i.e., sensors to enable reliable food safety monitoring throughout the whole future food supply chain. In addition, advanced monitoring options can help in the further optimization of the meat cultivation which may reduce the currently still high costs of production. This review presents an overview of the sensor monitoring options for the most relevant parameters of importance for meat cultivation. Examples of the various types of sensors that can potentially be used in CM production are provided and the options for their integration into bioreactors, as well as suggestions on further improvements and more advanced integration approaches. In favor of the multidisciplinary approach, we also include an overview of the bioreactor types, scaffolding options as well as imaging techniques relevant for CM research. Furthermore, we briefly present the current status of the CM research and related regulation, societal aspects and challenges to its upscaling and commercialization.

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Animal tissue culture principles and applications

Cited by 98 publications

References 46 publications

In vitro infection of Madin-Darby bovine kidney (MDBK) cells with Eimeria acervulina sporozoites: quantitative analysis of parasite cellular invasion and replication using real-time polymerase chain reaction (PCR)

In vitro infection of Madin-Darby bovine kidney (MDBK) cells with Eimeria acervulina sporozoites: quantitative analysis of parasite cellular invasion and replication using real-time polymerase chain reaction (PCR)

High Sensitivity and Low-Cost Flavin luciferase (FLUX)-based Reporter Gene for Mammalian Cell Expression

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production

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