293 CELLS HUMAN EMBRYO KIDNEY (C15-1310-345)

Cell Line Origin

Human Embryo Kidney

Cell Line Description

Transformed with sheared human Ad5 DNA. Sensitive to human adenoviruses and adenovirus DNA. Can be used to isolate transformation defective host-range mutants of Ad5 and for titrating human adenoviruses. This is a hypotriploid human cell line. The modal chromosome number was 64, occurring in 30% of cells. The rate of cells with higher ploidies was 4.2%. The der (1)t(1;15) (q42;q13), der(19)t(3;19)(q12;q13),der(12)t(8;12) (q22;p13) and four other marker chromosomes were common to most cells. Five other markers occurred in some cells only. The marker der(1) and M8 (or Xq+) were often paired. There were four copies of N17 and N22. Noticeably in addition to three copies of X chromosomes, there were paired Xq+ and a single Xp+ in most cells. The Ad insert was shown to consist of a colinear segment from nucleotides 1 to 4344 integrated into chromosome 19 (19q13.2). Expression of an unusual cell surface receptor for vitronectin has been reported. This is composed of the integrin β-1 subunit and the vitronectin receptor α-v subunit. Detach at room temperature; may take up to seven days to reattach.

Application

The human embryonic kidney 293 cell line has been used:

• to study the effects of the Bt insecticidal toxins Cry1Ab and Cry1Ac alone, or with a glyphosate-based herbicide.
• for toxicological studies of ethoxylated adjuvants of glyphosate-based herbicides.
• for stable expression of either type 1 or type 2 11-βHSD (11 β-hydroxysteroid dehydrogenase) after transfection with 11-βHSD cDNA.
• as a host to transfect TLR4 cDNA (HEK-TLR4) to express TLR4 mRNA and protein and induce IL-8 (interleukin-8) promoter activity in response to NE (neutrophil elastase).

Transformation studies

DNA Profile

STR-PCR Data: Amelogenin: X
CSF1PO: 11,12
D13S317: 12,14
D16S539: 9,13
D5S818: 8,9
D7S820: 11,12
THO1: 7,9.3
TPOX: 11
vWA: 16,19

Culture Medium

EMEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) + 10% FCS

Subculture Routine

Split sub-confluent cultures (70-80%) 1:2 to 1:6 i.e. seeding at 2-5x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37°C. When resuscitating a frozen ampoule of cells we recommend the cells are seeded at the higher seeding level i.e. 5 x10,000/cm². Cells may take up to 7 days to attach after resuscitation and subculture. Cells detach easily at room temperature or during transit, therefore growing cultures may be received with cells in suspension. In this event, centrifuge contents of flask and reseed to allow re-attachment of cells. These cells should be frozen in 5% DMSO : 95% FCS.

Other Notes

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biological source: human kidney (embryonic). growth mode: Adherent. karyotype: 2n 46, hypotriploid, modal no. 64. morphology: Epithelial. products: Not specified. receptors: Not specified. technique(s): cell culture | mammalian: suitable. shipped in: dry ice. storage temp.: −. 196°C.