The CD14 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, engineered for disruption of the CD14 gene. This product provides a heterogeneous pool of cells with targeted loss of CD14 expression, enabling robust studies of CD14-dependent innate immune signaling without clonal selection biases. As a polyclonal knockout model, it reflects the genetic diversity of the edited population while maintaining the parental epithelial characteristics, making it suitable for assays in immunology, cancer biology, and host-pathogen interactions.
The parental AGS cell line was originally established from a gastric adenocarcinoma of a 54-year-old female and serves as a well-characterized model of the gastric mucosal epithelium. These adherent epithelial cells retain key features of gastric mucosal cells, including responsiveness to microbial stimuli and inflammatory mediators. AGS cells have been widely used to investigate Helicobacter pylori infection mechanisms, gastric carcinogenesis, and mucosal innate immunity, providing a relevant backdrop for examining CD14-mediated processes in the stomach microenvironment.
CD14 is a glycosylphosphatidylinositol (GPI)-anchored co-receptor predominantly expressed on myeloid cells and mucosal epithelial surfaces. It binds bacterial lipopolysaccharide (LPS) in complex with LPS-binding protein (LBP) and transfers it to the Toll-like receptor 4 (TLR4)/MD-2 complex, initiating downstream signaling cascades. These include MyD88-dependent activation of interleukin-1 receptor-associated kinase 1 (IRAK1) and TRAF6, and TRIF-dependent pathways, both converging on NF-??B and AP-1 transcription factors. This leads to transcriptional induction of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-??), interleukin-6 (IL-6), and IL-1??, and production of type I interferons. CD14 thus functions as a critical gatekeeper for innate immune recognition of Gram-negative bacteria and other pathogen-associated molecular patterns, with its activity modulated by upstream stimuli including TNF-??, IL-1??, and LPS itself.
Disruption of CD14 in AGS cells creates a powerful loss-of-function model to dissect the specific role of this co-receptor in gastric epithelial innate immunity. Given the importance of CD14 in recognizing Helicobacter pylori LPS and mediating inflammatory responses, these knockout cells facilitate the study of how CD14 contributes to gastric mucosal inflammation and the development of gastric cancer. Researchers can directly assess CD14-dependent signaling versus alternative pattern-recognition receptors, explore cytokine production profiles, and evaluate the role of CD14 in epithelial barrier integrity and tumor microenvironment dynamics. Moreover, this model permits investigation of CD14 involvement in drug resistance mechanisms by comparing edited and parental cells under chemotherapeutic stress.
Typical experimental applications include LPS stimulation assays to quantify cytokine secretion via ELISA, NF-??B reporter assays to measure transcriptional activity, and flow cytometry to validate CD14 surface loss. The polyclonal population is suitable for western blotting and RT-qPCR to assess pathway components and phosphorylation. Proliferation, migration, and invasion assays further enable examination of CD14??s impact on gastric cancer cell behavior. This knockout model supports research into sepsis, inflammatory bowel disease, cardiovascular disease, and gastric cancer pathogenesis. For additional technical details, please contact Ascent Research.
