EML-3C細(xì)胞

生長狀態(tài)： 貼壁生長

運(yùn)輸方式： 凍存運(yùn)輸

器官來源： 外周血

ATCC Number： CRL-2996?

細(xì)胞形態(tài)： 其他

數(shù)量： 大量

年限： 2 years old

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 其他

Designations: EML-3C

Depositors: R Montelaro

EML-3C細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Equus caballus

Morphology: macrophage-like

Source: Organ: peripheral blood

Cell Type: macrophage-like

Breed: Portuguese autochthonous (Garrano)

Cellular Products: non-specific esterase

produces nitrites in response to LPS stimulation

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Isolation:EML-3C細(xì)胞 Isolation date: May 2005

limiting dilution

Applications: study of interaction between EIAV and macrophages; lentiviruses persistence; cytopathicity mechanisms

Receptors: equine infectious anemia virus (EIAV) cellular receptor, expressed

Virus Susceptibility: Equine infectious anemia virus

Antigen Expression: CZ2.2+, Ki-M6 (CD68)+, MHC-I+, MHC-II+

Age: 2 years old

Gender: male

Comments: The EML-3C cell line was established in 2005 from isolated peripheral blood mononuclear cells of a two-year-old male Portuguese Garrano horse. These cells possess functional properties of macrophages such as non-specific esterase (NSE) activity, are able to phagocytose fluorescent bioparticles and produce nitrites in response to lipopolysaccharide (LPS) stimulation. The EML-3C cell line expresses the EIA V receptor for cellular entry (ELR1) and supports replication of the virulent equine infectious anemia virus (EIA Vpv). This cell line can be used as a valuable tool for studying equine macrophage functions, lentivirus infection, and the equine immune system. [16173840]

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to 500 ml of the base medium:

fetal bovine serum (FBS) to a final concentration of 10%

horse serum (HS) to a final concentration of 10%

0.1 mM non-essential amino acids

EML-3C細(xì)胞Temperature: 37.0°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Subculturing: Protocol: Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37.0°C to facilitate dispersal.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 X 104 to 4 X 104 viable cells/cm2 is recommended.

Incubate cultures at 37.0°C.

Subcultivation ratio: EML-3C細(xì)胞A subcultivation ratio of 1:4 to 1:8 is recommended.

Medium renewal: Every 2 to 3 days

Preservation: Freeze medium: complete growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Related Products: Recommended medium (without the additional serum described under ATCC Medium): ATCC 30-2002

Recommended fetal bovine serum: ATCC 30-2020

Recommended horse serum: ATCC 30-2040

Trypsin EDTA Solution: ATCC 30-2101

Phosphate-buffered saline: ATCC 30-2200

Cell culture tested DMSO: ATCC 4-X

MEM Non-Essential Amino Acid Solution, 100x, ATCC 30-2116

References: 16173840: Isabel Fidalgo-Carvalho et al. Characterization of an equine macrophage cell line: application to studies of EIAV infection. Vetenary Microbiol. EML-3C細(xì)胞136 (1-2): 8?19, 2009. PubMed: 19038510