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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
Author(s) -
Saberi Samaneh,
Pournasr Behshad,
Farzaneh Zahra,
Esmaeili Maryam,
Hosseini Mahmoud Eshagh,
Baharvand Hossein,
Mohammadi Marjan
Publication year - 2018
Publication title -
helicobacter
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.206
H-Index - 79
eISSN - 1523-5378
pISSN - 1083-4389
DOI - 10.1111/hel.12489
Subject(s) - cytokeratin , immunostaining , helicobacter pylori , in vitro , embryonic stem cell , progenitor cell , biology , cell culture , microbiology and biotechnology , immunology , pathology , immunohistochemistry , stem cell , medicine , genetics , biochemistry , gene
Background Most two‐ dimensional in vitro models for studying host‐ H. pylori interactions rely on tumor‐derived cell lines, which harbor malignant alterations. The recent development of human gastric organoids has overcome this limitation and provides a highly sophisticated, yet costly, short‐term model for H. pylori infection, with restricted use in low‐budget centers. Method Tissue specimens from upper, middle, and lower stomachs of H. pylori ‐negative volunteers were collectively dispersed and cultured on mouse embryonic fibroblast ( MEF ) or collagen‐coated plates. Gastric primary cells ( GPC s) were evaluated by light microscopy, immunostaining, qRT ‐ PCR and ELISA analysis of cellular secretions, before and after H. pylori infection. Results The formation and long‐term (up to 1 year) maintenance of GPC s was highly dependent on adherent inactivated MEF cells, cultured in enriched media. These cells were multipassageable and able to undergo stable freezer storage and subsequent revival. The cellular composition of GPC s included the combination of cytokeratin 18 ( CK 18) and E‐cadherin (E‐cad)‐positive epithelial cells, MUC 5 AC ‐positive gastric cells, and leucine‐rich repeat containing G protein‐coupled receptor 5 ( LGR 5)‐positive progenitor cells. These cells produced significant amounts of gastric pepsinogens I and II . GPC s also allowed for extended (up to 96 hours) H. pylori infection, during which they underwent morphological alterations (cellular vacuolation and elongation) and hyperproduction of gastric pepsinogens and inflammatory cytokines ( IL ‐8 and TNF ‐α). Conclusion 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.