產(chǎn)品名稱 |
3D4/31 |
商品貨號(hào) |
B206451 |
Organism |
Sus scrofa, pig |
Tissue |
lung |
Cell Type |
macrophage (alveolar); immortalized with SV40 large T antigen |
Product Format |
frozen |
Morphology |
macrophage |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells contain SV40 viral DNA sequences]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
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  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
|
Storage Conditions |
liquid nitrogen vapor phase |
Images |
 |
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 . 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). |
Clinical Data |
unknown |
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 |
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. |
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%
|
Subculturing |
Volumes used in this protocol are for 75 cm2 flask; 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%Trypsin-0.53mM 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°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 103 to 2 x104 viable cells/cm2 is recommended.
- Place culture vessels in incubators at 37°C. Subculture when cell concentration is between 8x104 and 1 x 105 cells/cm2.
Subcultivation Ratio: 1:6 to 1:8 twice 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. |
Cryopreservation |
Freeze medium: Fetal bovine serum, 80%; complete growth medium, 10%; DMSO, 10% Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
Population Doubling Time |
19 hours |
Name of Depositor |
J Gren |
Year of Origin |
December, 1998 |
References |
also pseudorabies virus (PRV), classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), swine poxvirus, African swine fever virus (ASFV), herpes simplex virus (HSV), parainfluenza virus, bovine adenovirus (BAV), vaccinia virus (VV), and porcine adenovirus (PAV).
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
|