Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line

Kaiser, Bastian; Böttner, Martina; Wedel, Thilo; Brunner, Ronald M.; Goldammer, Tom; Lesko, Szilvia; Gäbel, Gotthold; Gleich, Alexander; Pfannkuche, Helga

doi:10.1159/000453394

Cited by 12 publications

(7 citation statements)

References 113 publications

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“…However, when such experiments focus on livestock, such as pigs, which are not only used for studying pig-specific diseases but also extensively as model for human intestinal pathophysiology [reviewed by 1 ], alternative models are still limited. Porcine cell lines, such as the small intestine-derived IPEC-J2 cells [ 2 ] and the colon-derived cell line PoCo83-3 [ 3 ] serve as a potential approach to investigate pathogen-host interactions [ 4 , 5 ]. However, these cell lines only contain enterocytes and lack other cell types such as goblet cells.…”

Section: Introductionmentioning

confidence: 99%

Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine

Hoffmann¹,

Schnepel²,

Langeheine³

et al. 2021

PLoS ONE

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Gastrointestinal infectious diseases remain an important issue for human and animal health. Investigations on gastrointestinal infectious diseases are classically performed in laboratory animals leading to the problem that species-specific models are scarcely available, especially when it comes to farm animals. The 3R principles of Russel and Burch were achieved using intestinal organoids of porcine jejunum. These organoids seem to be a promising tool to generate species-specific in vitro models of intestinal epithelium. 3D Organoids were grown in an extracellular matrix and characterized by qPCR. Organoids were also seeded on permeable filter supports in order to generate 2D epithelial monolayers. The organoid-based 2D monolayers were characterized morphologically and were investigated regarding their potential to study physiological transport properties and pathophysiological processes. They showed a monolayer structure containing different cell types. Moreover, their functional activity was demonstrated by their increasing transepithelial electrical resistance over 18 days and by an active glucose transport and chloride secretion. Furthermore, the organoid-based 2D monolayers were also confronted with cholera toxin derived from Vibrio cholerae as a proof of concept. Incubation with cholera toxin led to an increase of short-circuit current indicating an enhanced epithelial chloride secretion, which is a typical characteristic of cholera infections. Taken this together, our model allows the investigation of physiological and pathophysiological mechanisms focusing on the small intestine of pigs. This is in line with the 3R principle and allows the reduction of classical animal experiments.

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Section: Introductionmentioning

confidence: 99%

Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine

Hoffmann¹,

Schnepel²,

Langeheine³

et al. 2021

PLoS ONE

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“…The previously common method for cell immortalization is to introduce the simian virus 40 (SV40) T antigen into target primary cells. The large T antigen forms complexes with tumor suppressors, such as pRB-1 and p53, and induces DNA synthesis in both dividing and quiescent cells, leading to extension of the lifespan of the cells [26]. This approach has been used for transformation of various primary cell types [27][28][29].…”

Section: Discussionmentioning

confidence: 99%

Establishment of A Reversibly Inducible Porcine Granulosa Cell Line

Bai

Zhu

Mao

et al. 2020

Cells

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Granulosa cells (GCs) are the key components of ovarian follicles for regulating oocyte maturation. Previous established GC lines have allowed prolonged proliferation, but lost some physiological features owing to long-term immortalization. This study was to establish an induced immortal porcine GC line with reversible proliferation status by the tetracycline inducible (Tet-on) 3G system. Our conditional immortal porcine GCs (CIPGCs) line steadily propagated for at least six months and displayed primary GC morphology when cultured in the presence of 50 ng/mL doxycycline [Dox (+)]. Upon Dox withdrawal [Dox (–)], Large T-antigen expression, reflected by mCherry fluorescence, gradually became undetectable within 48 h, accompanied by less proliferation and size increase. The levels of estradiol and progesterone, and the expression of genes associated with steroid production, such as CYP11A1 (cytochrome P450 family 11), 3β-HSD (3β-hydroxysteroid dehydrogenase), StAR (steroidogenic acute regulatory protein), and CYP19A1 (cytochrome P450 family 19 subfamily a member 1), were all significantly higher in the Dox (–) group than Dox (+) group. The CIPGCs could switch into a proliferative state upon Dox induction. Interestingly, the expression of StAR and CYP19A1 in the CIPGCs (–Dox) was significantly increased by adding porcine follicular fluid (PFF) to mimic an ovary follicle environment. Moreover, PFF priming the CIPGCs in Dox (–) group resulted in similar estradiol production as that of primary GC, and enabled this cell line to respond to gonadotrophins in estradiol production. Collectively, we have established an inducible immortal porcine GC line, which offers a unique and valuable model for future research on the regulation of ovarian functions.

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“…In 2017 Kaiser et al ( 37 ) established a new porcine colonic cell line PoCo83-3. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen.…”

Section: In Vitro Cell-based Modelsmentioning

confidence: 99%

Assessing Intestinal Health. In Vitro and Ex vivo Gut Barrier Models of Farm Animals: Benefits and Limitations

Ghiselli

Rossi²,

Piva

et al. 2021

Front. Vet. Sci.

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Animal performance is determined by the functionality and health of the gastrointestinal tract (GIT). Complex mechanisms and interactions are involved in the regulation of GIT functionality and health. The understanding of these relationships could be crucial for developing strategies to improve animal production yields. The concept of “gut health" is not well defined, but this concept has begun to play a very important role in the field of animal science. However, a clear definition of GIT health and the means by which to measure it are lacking. In vitro and ex vivo models can facilitate these studies, creating well-controlled and repeatable conditions to understand how to improve animal gut health. Over the years, several models have been developed and used to study the beneficial or pathogenic relationships between the GIT and the external environment. This review aims to describe the most commonly used animals' in vitro or ex vivo models and techniques that are useful for better understanding the intestinal health of production animals, elucidating their benefits and limitations.

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Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line

Cited by 12 publications

References 113 publications

Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine

Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine

Establishment of A Reversibly Inducible Porcine Granulosa Cell Line

Assessing Intestinal Health. In Vitro and Ex vivo Gut Barrier Models of Farm Animals: Benefits and Limitations

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