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3D4/31
3D4/31
規(guī)格:
價(jià)格:
編號(hào):B206451
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱
3D4/31
商品貨號(hào)
B206451
Organism
Sus scrofa, pig
Tissue
lung
培養(yǎng)基
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. RefWeingartl 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.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25%Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. 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.
  6. 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

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