品系： Landrace
ATCC Number： CRL-2843?
細(xì)胞形態(tài)： 單核細(xì)胞/巨噬細(xì)胞
生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)
年限： 27 days
運(yùn)輸方式： 凍存運(yùn)輸
數(shù)量： 大量
器官來源： 肺
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： 豬
Designations: 3D4/21
3D4/21細(xì)胞Depositors: J Gren
Biosafety Level: 2 [cells containing SV40 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Sus scrofa
Morphology: macrophage
Source: Organ: lung
Strain: Landrace
Cell Type: macrophage macrophage (alveolar); immortalized with SV40 large T antigentransformed with pSV3-neo
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Isolation: Isolation date: December, 1998
Applications: These porcine myelomonocytic cell lines may have a wide variety of applications in porcine virology and immunology [PubMed:12088830].
3D4/21細(xì)胞Clone 3D4/21 can produce Bovine adenovirus type 3 (BAV-3) to markedly higher titers than clones 3D4/2 and 3D4/31. Addition of DMSO improved the capability of clone 3D4/21 to replicate the field isolate of African swine fever virus (ASFV/Lillie) compared to the other clones.
Single cell cloning and selection in G-418 of the 3D4 parental cell line resulted in establishment of 3D4/2 (ATCC CRL-2845), 3D4/21 (ATCC CRL-2843) and 3D4/31 (ATCC CRL-2844).
The parental porcine monomyeloid cell line, 3D4, was established in December of 1998 following transfection of primary porcine alveolar macrophage cultures with the pSV3neo plasmid.
A subpopulation of each cell line was positive, to varying degrees depending on the media formulation, for nonspecific esterase activity and phagocytosis.
Virus Susceptibility: Bovine adenovirus 3
Classical swine fever virus
Human parainfluenza virus 3
Swinepox virus
Vesicular stomatitis New Jersey virus
Porcine adenovirus
Herpes simplex virus 1
African swine fever virus
Pseudorabies virus
Vaccinia virus
Swine vesicular disease virus
Age: 27 days
Gender: unknown
Comments: 3D4/21細(xì)胞The parental porcine monomyeloid cell line, 3D4, was established in December of 1998 following transfection of primary porcine alveolar macrophage cultures with the pSV3neo plasmid. The plasmid carries the genes for neomycin resistance and SV40 large T antigen. Single cell cloning and selection in G-418 of the 3D4 parental cell line resulted in establishment of 3D4/2 (ATCC CRL-2845), 3D4/21 (ATCC CRL-2843) and 3D4/31 (ATCC CRL-2844). A subpopulation of each cell line was positive, to varying degrees depending on the media formulation, for nonspecific esterase activity and phagocytosis. Clone 3D4/21 can produce Bovine adenovirus type 3 (BAV-3) to markedly higher titers than clones 3D4/2 and 3D4/31. Addition of DMSO improved the capability of clone 3D4/21 to replicate the field isolate of African swine fever virus (ASFV/Lillie) compared to the other clones. These porcine myelomonocytic cell lines may have a wide variety of applications in porcine virology and immunology [PubMed:12088830].
Propagation: ATCC complete growth medium: RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1.0 mM sodium pyruvate supplemented with 0.1 mM nonessential amino acids, 90%; fetal bovine serum, 10%
Atmosphere: 5% CO2 in air recommended
Temperature: 37.0°C
Subculturing: Protocol: Volumes used in this protocol are for 75cm2 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 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that 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?C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.3D4/21細(xì)胞 An inoculum of 5 X 10(3) to 7 X 10(3) viable cells/cm2 is recommended.
Incubate cultures at 37?C. Subculture when cell concentration reaches between 3 X 10(5) and 4 X 10(5) cells/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Two to three times weekly
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: about 18 hrs
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
Erythrosin B vital stain solution:ATCC 30-2404
Trypan Blue vital stain solution:ATCC 30-2402
MEM Non-Essential Amino Acid Solution, 100x:ATCC 30-2116
References: 92770: Weingartl HM, et al. 3D4/21細(xì)胞Continuous porcine cell lines developed from alveolar macrophages: partial characterization and virus susceptibility. J. Virol. Methods 104: 203-216, 2002. PubMed: 12088830
