Cell line:

FLK-BLV-044

NTCC no.:

ACC 153

Species validation:

ovine (sheep) (Ovis aries)

Cell type:

embryonal kidney cells

Origin:

established from the embryonal kidney cells of an ovine in 1976; cells are susceptible to infection with BLV (bovine leukemia virus) leading to the formation of syncythia

Reference(s):

not published

Biosafety level:

1

Risk assessment:

The cell line is infected with bovine leukemia virus (BLV) and produces active viruses. The viruses are not pathogenic to humans, but may be pathogenic to animals.

NTCC Cell Culture Data:

Morphology:

large fibroblast-like cells growing as monolayers;

Medium:

90% MEM (with Earle's salts) + 10% h.i. FBS optional: + 850 mg/L NaHCO3 + 120 mg/L sodium pyruvate + 1x non-essential amino acids

Subculture:

seed out about 1-2 x 106 cells/80 cm2; split confluent culture 1:2 to 1:8 every 3-5 days using trypsin/EDTA

Incubation:

at 37 °C with 5% CO2

Doubling time:

ca. 50 hours

Harvest:

cell harvest of about 4 x 106/80 cm2

Storage:

frozen with 82.5% medium, 10% FBS, 7.5% DMSO

NTCC Scientific Data:

Mycoplasma:

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

Species validation:

Species-level identification was carried out by mitochondrial Cytochrome C Oxidase Subunit 1 (COI) DNA barcoding according standard ANSI/ATCC ASN-0003-2015 and revealed Ovis aries species.

Viruses:

ELISA: reverse transcriptase positive; PCR: SMRV -

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