The CD14 Knockout 143B Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 143B human osteosarcoma cell line, with targeted disruption of the CD14 gene. This product provides a heterogeneous pool of cells carrying diverse loss-of-function edits, enabling functional studies of CD14-dependent pathways without clonal selection. The polyclonal format preserves genetic diversity while abolishing CD14-mediated signaling, offering a robust model for investigating innate immune responses in a bone cancer context.
The 143B parental line is a highly tumorigenic and metastatic osteosarcoma cell line originally derived from the HOS family. It is widely employed as a model system for studying bone cancer progression, metastasis mechanisms, and the tumor microenvironment. 143B cells exhibit characteristic features of advanced osteosarcoma, including rapid proliferation and the capacity to form osteolytic lesions in vivo, making them a relevant host for examining interactions between inflammatory signaling and osteosarcoma biology.
CD14 operates as a glycosylphosphatidylinositol-anchored co-receptor that binds bacterial lipopolysaccharide (LPS) in concert with the serum protein LBP and presents it to the TLR4/MD-2 complex. This engagement initiates intracellular cascades mediated by adaptors MyD88 and IRAK, leading to TRAF6-dependent activation of the IKK complex and subsequent nuclear translocation of NF-?B, as well as stimulation of MAP kinases including ERK, JNK, and p38. These pathways converge on the transcriptional upregulation of proinflammatory cytokines such as TNF-??, IL-6, and IL-1??, establishing a positive feedback loop driven by TNF-?? and IL-1?? themselves. Consequently, CD14 is central to innate immune recognition and amplification of inflammatory responses.
In the context of 143B osteosarcoma cells, CD14 knockout allows dissection of innate immune signaling contributions to tumor cell behavior and the metastatic niche. Osteosarcoma cells can respond to microbial-derived products and endogenous danger signals within the bone microenvironment, potentially influencing tumor growth, invasion, and immune evasion. Disruption of CD14 in this lineage provides a means to evaluate the role of TLR4-mediated inflammation in osteosarcoma pathophysiology, including interactions with osteoclasts and immune infiltrates that shape disease progression.
Researchers can employ this knockout model to investigate LPS-induced signal transduction, NF-?B activation, and cytokine production using techniques such as LPS stimulation assays, NF-?B reporter assays, ELISA quantitation of secreted TNF-?? or IL-6, and western blotting for phosphorylated MAP kinases. Additional applications include phagocytosis studies, drug screening for anti-inflammatory compounds, and co-culture experiments mimicking the tumor microenvironment. The polyclonal population is suited for functional genomics screens and validation of CD14-dependent metastatic phenotypes. For further details and ordering information, please contact Ascent Research.
