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Cells contain Adenovirus DNA sequences
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The HEK293S GnTIˉ cell line was established by methanesulfonate mutagenesis followed by Ricin selection. HEK293S GnTIˉ cells do not have N-acetyl-glucosaminyltransferase I (GnTI) activity and therefore lack complex N-glycans.RefReeves P, et al. Structure and function in rhodopsin: High-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. Proc. Natl. Acad. Sci. USA 99 (21): 13419-13424, 2002. PubMed: 12370423 This cell line has been successfully used to overexpress a wide variety of mammalian membrane proteins. There were two versions of HEK293S GnTIˉ cells in the original publication. One was HEK293S GnTIˉ (ATCC CRL-3022), and the other version was HEK293S GnTIˉ variant constructed with tetracycline-inducible system. CRL-3022 does not contain tetracycline-inducible system or pcDNA6-TR vector.
Note: After thawing, these cells take 4 to 5 days to begin attaching to the flask. Perform first medium replacement at that time. The cells will grow in chains or small clumps that will eventually attach and spread. Attachment can be enhanced by using Corning CellBIND® flasks.
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Reeves P, et al. Structure and function in rhodopsin: High-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. Proc. Natl. Acad. Sci. USA 99 (21): 13419-13424, 2002. PubMed: 12370423
Graham FL, et al. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 36: 59-72, 1977. PubMed: 886304
Graham FL, et al. Defective transforming capacity of adenovirus type 5 host-range mutants. Virology 86: 10-21, 1978. PubMed: 664220