Helicobacter. 2018;e12489. wileyonlinelibrary.com/journal/hel | 1 of 12 https://doi.org/10.1111/hel.12489 © 2018 John Wiley & Sons Ltd DOI: 10.1111/hel.12489 ORIGINAL ARTICLE A simple and cost-efficient adherent culture platform for human gastric primary cells, as an in vitro model for Helicobacter pylori infection Samaneh Saberi 1 | Behshad Pournasr 2 | Zahra Farzaneh 2 | Maryam Esmaeili 1 | Mahmoud Eshagh Hosseini 3 | Hossein Baharvand 2,4 | Marjan Mohammadi 1 1 HPGC Research Group, Department of Medical Biotechnology, Pasteur Institute of Iran, Tehran, Iran 2 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran 3 Gastroenterology Department, Amiralam Hospital, Tehran University of Medical Sciences, Tehran, Iran 4 Department of Developmental Biology, University of Science and Culture, Tehran, Iran Correspondence Marjan Mohammadi, HPGC Research Group, Department of Medical Biotechnology, Pasteur Institute of Iran, Tehran, Iran. Email: marjan.mohammadi@pasteur.ac.ir and Hossein Baharvand, Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Email: baharvand@royaninstitute.org Funding information Pasteur Institute of Iran: Grant/Award Number: #722; and #TP-9121 and Royan Institute: Grant/Award Number: 94000090 Abstract Background: Most two- dimensional in vitro models for studying host- H. pylori inter- actions rely on tumor-derived cell lines, which harbor malignant alterations. The re- cent 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 (GPCs) were evalu- ated 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 GPCs 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 GPCs included the combination of cytokeratin 18 (CK18) and E-cadherin (E-cad)-positive epithelial cells, MUC5AC-positive gastric cells, and leucine-rich repeat containing G protein-coupled receptor 5 (LGR5)-positive pro- genitor cells. These cells produced significant amounts of gastric pepsinogens I and II. GPCs also allowed for extended (up to 96 hours) H. pylori infection, during which they underwent morphological alterations (cellular vacuolation and elongation) and hyper- production 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. KEYWORDS epithelial, gastric, Helicobacter pylori, human, primary, progenitor 1 | INTRODUCTION The gastric lumen, in the gastrointestinal tract, remains a se- cluded entity, which has made its study difficult for interested investigators. On the other hand, the mere fact that a previously believed sterile environment can become colonized with a carcino- genic pathogen (Helicobacter pylori ), has attracted extensive inter- est in studying the real-time molecular details of its host-pathogen