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| General Information | |
| Organism | Sus scrofa, pig |
| Cell Line Description | 3D4/21 is an alveolar cell line isolated from 27-day-old pig lungs in 1998. The 3D4/21 cell line is derived from a single-cell clone of the parent (continuous porcine alveolar macrophage cell line). This cell is an effective tool for studying the immune characteristics and viral infection mechanisms of pigs. There are 844 InDels greater than 1 kb in the 3D4/21 cell genome, of which 12 regions are deletions greater than 100 kb. In addition, compared with primary porcine alveolar macrophages, 82 genes (including CD163) were transcribed missing in 3D4/21 cells, and 72 genes were transcribed. Further combined with Hi-C structure, it was found that the topologically associated domain (TAD) fusion caused by the deletion may lead to abnormal gene function. |
| Tissue | Lung |
| Morphology | Macrophage |
| Gender | Unknown |
| Age | 27 days |
| Product Format | Frozen |
| Growth Mode | Adherent |
| Biosafety Level | 2 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. |
| Applications | 1. Viral pathogenesis and vaccine development 2. Innate immunity studies 3. Gene expression and regulation 4. Drug screening and development |
| Shipped In | Dry ice |
| Storage Temperature | −196°C |
| Additional Info | It has been reported that 3D4/21 can support the growth of cell-adapted ASFV-Lisbon 61 and field isolate Lillie SI/85, but it was unable to maintain replication of the genotype II ASFV-HLJ/18 strain. However, a more recent study showed that the ASFV genotype II strain CN/GS/2018 can attach and enter 3D4/21 cells, resulting in genome replication rate of up to 10 6 copies/mL. |
| Characteristics | |
| Gene Expressed | SV40 large T antigen transformed with pSV3-neo |
| Virus Susceptibility | Bovine adenovirus 3, Classical swine fever virus, Human parainfluenza virus 3, Swinepox virus, Herpes simplex virus 1, African swine fever virus, Vesicular stomatitis New Jersey virus, Vaccinia virus, Porcine adenovirus, Pseudorabies virus, Swine vesicular disease virus |
| Tumorigenic | No |
| Mycoplasma Test | Negative |
| Culture Conditions and Handling | |
| Subculturing | The volumes used in this protocol are for 75cm2 flasks. For culture vessels of other sizes, the amount of dissociation medium should be proportionally reduced or increased. 1. Remove and discard the culture medium. 2. Briefly rinse the cell layer with 0.25% (w/v) trypsin - 0.53 mM EDTA solution to remove all traces of serum containing trypsin inhibitors. 3. Add 2.0 to 3.0 mL of trypsin-EDTA solution to the flask and observe the cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping, do not agitate the cells by tapping or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach can be placed at 37℃ to facilitate dispersion. 4. Add 6.0 to 8.0 mL of complete growth medium and gently pipette to aspirate the cells. 5. Add an appropriate amount of the cell suspension to a new culture vessel. The recommended inoculum size is 5 x 103 to 7 x 103 viable cells/cm2. 6. Incubate the culture at 37℃. Subculture when the cell concentration reaches between 3 x 105 and 4 x 105 cells/cm2. |
| Medium Renewal | 2 to 3 times weekly |
| Subcultivation Ratio | The ratio of 1:6 to 1:8 is recommended. |
| Culture 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% |
| Culture Conditions | Atmosphere: Air, 95%; CO2, 5%; Temperature: 37℃ |
| Cryopreservation | Complete growth medium supplemented with 5% (v/v) DMSO |
The above is only part of a part of cell line products. If you don’t find the cell line you want, Creative Biogene can also provide stable cell line generation service with the best prices and fastest turnaround time for you! Contact us for more information or to request a quote.
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|---|---|---|
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