Cell line:

293

NTCC no.:

ACC 305

Species:

human (Homo sapiens)

Cell type:

embryonal kidney

Origin:

established from a human primary embryonal kidney transformed by adenovirus type 5 (Ad 5); classified as risk category 1 according to the German Central Commission for Biological Safety (ZKBS); cell line also known as HEK-293 (human embryonic kidney-293)

Reference(s):

14573

Depositor:

Dr. W. Dirks, NTCC, Braunschweig, Germany

Biosafety level:

1, GMO-S1

Risk assessment:

The cell line was established by transfection of the cells with sheared DNA of adenovirus type 5. The cells contain 4 to 5 copies of the left end of the virus (12% of the viral genome) including E1a and E1b genes and one copy of the right end of the virus (10% of the genome) including the E4 gene. No active viruses are produced.

Permissions and restrictions:

A, D

NTCC Cell Culture Data:

Morphology:

adherent fibroblastoid cells growing as monolayer

Medium:

90-95% Dulbeccos MEM + 5-10% h.i. FBS

Subculture:

split confluent culture 1:5 to 1:6 every 2-3 days (without using trypsin/EDTA, cells can be detached by tapping the flask); seed out at about 2-3 x 10⁶ cells/25 cm²

Incubation:

at 37 °C with 10% CO₂

Doubling time:

about 24-30 hours

Harvest:

cell harvest of about 40 x 10⁶ cells/175 cm²

Storage:

frozen with 70% medium, 20% FBS, 10% DMSO at about 1 x 10⁶cells/ampoule

NTCC Scientific Data:

Mycoplasma:

negative in DAPI, microbiological culture, RNA hybridization, PCR assays

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