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3D4/21 (ATCC
® CRL-2843Organism | Sus scrofa, pig |
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Tissue | lung |
Cell Type | macrophage macrophage (alveolar); immortalized with SV40 large T antigen transformed with pSV3-neo |
Product Format | frozen |
Morphology | macrophage |
Culture Properties | adherent |
Biosafety Level | 2 [Cells contain SV40 viral DNA sequences] |
Age | 27 days |
Gender | unknown |
Strain | Landrace |
Applications | These porcine myelomonocytic cell lines may have a wide variety of applications in porcine virology and immunology Ref. 3D4/21 猪肺泡巨噬细胞 |
Storage Conditions | liquid nitrogen vapor phase |
Derivation | 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. 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). |
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Virus Susceptibility | Bovine adenovirus 3 Classical swine fever virus , 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 |
Comments | The plasmid carries the genes for neomycin resistance and SV40 large T antigen. A subpopulation of each cell line (3D4/2 (ATCC CRL-2845), 3D4/21 (ATCC CRL-2843) and 3D4/31 (ATCC CRL-2844)) 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. |
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% |
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Subculturing | Volumes used in this protocol are for 75cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subc*tion Ratio: A subc*tion ratio of 1:6 to 1:8 is recommended Medium Renewal: Two to three times weekly Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
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Cryopreservation | Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC® Catalog No. 4-X. |
Culture Conditions | Temperature: 37°C Atmosphere: 5% CO2 in air recommended |
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