A simple and cost‐efficient adherent culture platform for human gastric primary cells, as an in vitro model for <i>Helicobacter pylori</i> infection

Saberi, Samaneh; Pournasr, Behshad; Farzaneh, Zahra; Hosseini, Mojtaba; Baharvand, Hossein; Mohammadi, Marjan

doi:10.1111/hel.12489

Cited by 4 publications

(10 citation statements)

References 66 publications

(171 reference statements)

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“…But there has been no mature cell culture system representing normal gastric cells at present. This problem has been paid attention to and some scientists have begun to investigate it specially and technically (Saberi et al , 2018). It is expectable in the future, a simple, consistent, and mature cell culture system representing normal gastric cells can be used to better understand H. pylori -induced gastric carcinogenesis.…”

Section: Discussionmentioning

confidence: 99%

Helicobacter pylori-Induced DNA Damage Is a Potential Driver for Human Gastric Cancer AGS Cells

Shi

Wang

Yanlei

et al. 2019

DNA and Cell Biology

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Helicobacter pylori is a major cause of gastric cancer. This study was aimed to explore the characteristic of DNA damage induced by H. pylori infection in gastric cancer AGS cells. After infection with H. pylori, the reactive oxygen species (ROS) levels in AGS cells were significantly higher than those in the uninfected cells. Cells with longer comet tails were detected after infection with H. pylori. The number of apurinic/apyrimidinic endonuclease 1- and phosphorylated H2AX-positive cells was significantly increased compared with the number of negative control cells. The expression of pChk1 and pChk2 was significantly upregulated by H. pylori infection. Cell growth was inhibited after H. pylori infection. All these results were dose dependent. The cell alterations were more significant upon infection with H. pylori at a multiplicity of infection (MOI) of 100:1 than at an MOI of 50:1. H. pylori infection can induce DNA single-strand breaks, DNA double-strand breaks, and cell cycle checkpoint activation after ROS generation in the gastric cancer cell line AGS, which is a potential driver for gastric cancer.

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Section: Discussionmentioning

confidence: 99%

Helicobacter pylori-Induced DNA Damage Is a Potential Driver for Human Gastric Cancer AGS Cells

Shi

Wang

Yanlei

et al. 2019

DNA and Cell Biology

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“…First and foremost, to understand the essence of the studied cells, we examined their gene expression and clustered them by drawing gene expression charts and heat maps. Of these cells, the nontransformed (non-cancerous) primary cells (GPCs 29 ) clustered with the normal gastric tissue. Whereas, the essence of MKN45 and AGS cells was closer together and clustered with the intestinal type gastric tumor and duodenal tissue.…”

Section: Discussionmentioning

confidence: 99%

“…2B), 3). At 24h PI, IL-8 production was detected in GPCs (529.8 ± 5.4 pg/10 6 cells) 29 , MKN45 (883.5 ± 12.7 pg/10 6 cells) and AGS (302.2 ± 9.7 pg/10 6 cells), as compared to uninfected controls (P < 0.0001, Supplementary Fig. 2C).…”

Section: Characterization Of H Pylori Coculturementioning

confidence: 93%

Infection with a hypervirulent strain of Helicobacter pylori primes gastric cells toward intestinal transdifferentiation

Saberi

Tashakoripour

Hosseini

et al. 2022

Microbial Pathogenesis

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Background: Intestinal metaplasia, gastric-to-intestinal transdifferentiation, occurs as a result of the misexpression of certain regulatory factors, leading to genetic reprogramming. Here, we have evaluated the H. pylori-induced expression patterns of these candidate genes.Methods: The expression levels of 1)tissue-speci c transcription factors (RUNX3, KLF5, SOX2, SALL4, CDX1 and CDX2), 2)stemness factors (TNFRSF19, LGR5, VIL1) and 3)tissue-speci c mucins (MUC5AC, MUC2) were evaluated by quantitative real-time PCR in gastric primary cells (GPCs), in parallel with two gastric cancer (MKN45 and AGS) cell lines, up to 96h following H. pylori infection.Results: Following H. pylori infection of GPCs, RUNX3 declined at 24h post infection (PI) (-6.2±0.3) and remained downregulated for up to 96h. Subsequently, overexpression of self-renewal and pluripotency transcription factors, KLF5 (3.6±0.2), SOX2 (7.6±0.5) and SALL4 (4.3±0.2) occurred. The expression of TNFRSF19 and LGR5, demonstrated opposing trends, with an early rise of the former (4.5±0.3) at 8h, and a simultaneous fall of the latter (-1.8±0.5). This trend was reversed at 96h, with the decline in TNFRSF19 (-5.5±0.2), and escalation of LGR5 (2.6±0.2) and VIL1 (1.8±0.3). Ultimately, CDX1 and CDX2 were upregulated by 1.9 and 4.7-fold, respectively. The above scenario was, variably observed in MKN45 and AGS cells. Conclusion: Our data suggests an interdependent gene regulatory network, induced by H. pylori infection. This interaction begins with the downregulation of RUNX3, upregulation of self-renewal and pluripotency transcription factors, KLF5, SOX2 and SALL4, leading to the downregulation of TNFRSF19, upregulation of LGR5 and aberrant expression of intestine-speci c transcription factors, potentially facilitating the process of gastric-to-intestinal transdifferentiation.

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“…The tissue specimens were minced, enzymatically digested and used for gastric primary culture on mouse embryonic broblast (MEF). GPCs were expanded in advanced DMEM/F12 (Gibco, USA), supplemented with growth factors and small molecules (2-dimensional medium) 29,104 An H. pylori (C142LD) strain, originally isolated from a gastric cancer patient, was selected for this study.…”

Section: Cell Culture and Tissue Specimensmentioning

confidence: 99%

Infection with a Hypervirulent Strain of Helicobacter Pylori Primes Gastric Cells Toward Intestinal Transdifferentiation

Saberi

Esmaeili

Tashakoripour

et al. 2021

Preprint

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Background: Intestinal metaplasia, gastric-to-intestinal transdifferentiation, occurs as a result of the misexpression of certain regulatory factors, leading to genetic reprogramming. Here, we have evaluated the H. pylori-induced expression patterns of these candidate genes.Methods: The expression levels of 1)tissue-specific transcription factors (RUNX3, KLF5, SOX2, SALL4, CDX1 and CDX2), 2)stemness factors (TNFRSF19, LGR5, VIL1) and 3)tissue-specific mucins (MUC5AC, MUC2) were evaluated by quantitative real-time PCR in gastric primary cells (GPCs), in parallel with two gastric cancer (MKN45 and AGS) cell lines, up to 96h following H. pylori infection. Results: Following H. pylori infection of GPCs, RUNX3 declined at 24h post infection (PI) (-6.2±0.3) and remained downregulated for up to 96h. Subsequently, overexpression of self-renewal and pluripotency transcription factors, KLF5 (3.6±0.2), SOX2 (7.6±0.5) and SALL4 (4.3±0.2) occurred. The expression of TNFRSF19 and LGR5, demonstrated opposing trends, with an early rise of the former (4.5±0.3) at 8h, and a simultaneous fall of the latter (-1.8±0.5). This trend was reversed at 96h, with the decline in TNFRSF19 (-5.5±0.2), and escalation of LGR5 (2.6±0.2) and VIL1 (1.8±0.3). Ultimately, CDX1 and CDX2 were upregulated by 1.9 and 4.7-fold, respectively. The above scenario was, variably observed in MKN45 and AGS cells.Conclusion: Our data suggests an interdependent gene regulatory network, induced by H. pylori infection. This interaction begins with the downregulation of RUNX3, upregulation of self-renewal and pluripotency transcription factors, KLF5, SOX2 and SALL4, leading to the downregulation of TNFRSF19, upregulation of LGR5 and aberrant expression of intestine-specific transcription factors, potentially facilitating the process of gastric-to-intestinal transdifferentiation.

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A simple and cost‐efficient adherent culture platform for human gastric primary cells, as an in vitro model for Helicobacter pylori infection

Abstract: We, hereby, present a simple, consistent, and cost-efficient gastric cell culture system, which provides a suitable model for extended in vitro infection of H. pylori. This platform can be employed for a variety of gastric-related research.

Cited by 4 publications

References 66 publications

Helicobacter pylori-Induced DNA Damage Is a Potential Driver for Human Gastric Cancer AGS Cells

Helicobacter pylori-Induced DNA Damage Is a Potential Driver for Human Gastric Cancer AGS Cells

Infection with a hypervirulent strain of Helicobacter pylori primes gastric cells toward intestinal transdifferentiation

Infection with a Hypervirulent Strain of Helicobacter Pylori Primes Gastric Cells Toward Intestinal Transdifferentiation

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