Jurkat JE6.1 NF-kB::eGFP Cell Line

Contamination with bacterial components is a potential issue with the biologics for research- and therapeutic uses, as they can activate the so-called pattern-recognition receptors in mammalian cells and trigger immune responses, clouding the experimental results or even causing health hazards. Such responses are often mediated by the Toll-like Receptors (TLRs), which recognize bacterial molecular patterns to activate, among other things, cytokine expression through the NF-κB pathway.

The current assessment method for bacterial contamination measures lipopolysaccharide (LPS), the active component of endotoxins. Endotoxin measurement relies on its reaction with the limulus amoebocyte lysate (LAL), derived from the horseshoe crab blood.3 While the method is well established, however, the use of primary lysate obtained from wild animals presents a major sustainability issue.4 Recombinant alternatives to the primary LAL method have recently been gaining acceptance, but all LAL-based methods measure only the endotoxin contamination, and other classes of bacterial components go undetected.

To address these unmet needs, the junket JE6.1 cell line has been engineered to form a group of cell lines, each expressing a specific set of TLRs and harboring an eGFP expression cassette fused to the NF-κB Response Element. The result is that each of these strains expresses eGFP in response to a specific class of bacterial molecules.

SCC635, Jurkat JE6.1 NF-κB::eGFP cell line, expresses only the endogenous TLR6 on its surface and is designed to serve as the negative control for the group. It is not responsive to any bacterial components tested thus far, including the LPS, (ligands for SCC630, the TLR4 cell line), the Gram positive- and Gram negative flagellins (ligands for SCC631, the TLR5 cell line), and FSL-1 (ligand for SCC632, the TLR2/TLR6 cell line), and expresses eGFP only when the NF-κB Response Element is chemically activated. It has also been tested negative for PAM3CSK4 (ligand for TLR2/TLR1).

Source
SCC635 cell line was created by the deletion of the TLR5 gene in the Jurkat JE6.1 cell line harboring the eGFP expression cassette fused to the NF-kB Response Element.

Jurkat JE6.1 NF-ĸB::eGFP cell line expresses only the endogenous TLR6 on its surface and is designed to serve as the negative control and is not responsive to any bacterial components.

Jurkat JE6.1 NF-kB::eGFP cells should be stored in liquid nitrogen. The cells can be cultured for at least 10 passages without significantly affecting cell marker expression and function.

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