运输方式： 冻存运输
细胞形态： 单核细胞/巨噬细胞
年限： 27 days
细胞类型： 其他细胞类型
是否是肿瘤细胞： 0
物种来源： 猪
数量： 大量
生长状态： 贴壁生长
器官来源： 肺
品系： Landrace
ATCC Number： CRL-2844™
Designations: 3D4/31
Depositors: J Gren
3D4/31细胞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
Virus Susceptibility: 3D4/31细胞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: 3D4/31细胞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. Addition of DMSO to clone 3D4/31 upregulates the expression of CD14 monocyte marker. 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: air, 95%; carbon dioxide (CO2), 5%
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. An inoculum of 2 X 10(3) to 2 X 10(4) viable cells/cm2 is recommended.
Incubate cultures at 37�C. Subculture when cell concentration reaches between 8 X 10(4) and 1 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: Fetal bovine serum, 80%; complete growth medium, 10%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 19 hours
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) 3D4/31细胞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. Continuous porcine cell lines developed from alveolar macrophages: partial characterization and virus susceptibility. J. Virol. Methods 104: 203-216, 2002. PubMed: 12088830
