Cat.No. : LVIM003Z
Titer: ≥1*10^7 TU/mL / ≥1*10^8 TU/mL / ≥1*10^9 TU/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | LVIM003Z |
| Description | Lentivirus expressing untagged SV40 Large T Antigen under the control of CMV promoter. This lentivirus contains no antibiotic selection marker. |
| Target Gene | Large T Antigen |
| Species | SV40 |
| Titer | Varies lot by lot, for example, ≥1*10^7 TU/mL, ≥1*10^8 TU/mL, ≥1*10^9 TU/mL etc. |
| Size | Varies lot by lot, for example, 100 ul, 500 ul, 1 mL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality lentivirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between lentivirus particle lots. | |
| Mycoplasma | Creative Biogene routinely tests for mycoplasma contamination using a mycoplasma detection kit. Cell lines are maintained for approximately 20 passages before being discarded and replaced with a new vial of early passage cells. Approximately 2 weeks after thawing, cell culture supernatants are tested for mycoplasma contamination. Creative Biogene ensures that lentiviral products are free of mycoplasma contamination. |
| Purity | Creative Biogene evaluates the level of impurities, such as residual host cell DNA or proteins, in prepared lentiviral vectors to ensure they meet quality standards. |
| Sterility | The lentiviral samples were inoculated into cell culture medium for about 5 days and the growth of bacteria and fungi was tested. Creative Biogene ensures that the lentiviral products are free of microbial contamination. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of lentivirus to deliver genetic material into target cells, and assess gene expression and functional activities. |
| Proviral Identity Confirmation | All Creative Biogene lentiviral vectors are confirmed to have correctly integrated provirus using PCR. This test involves transducing cells with serial dilutions of the lentiviral vector, harvesting the cells a few days later, and isolating genomic DNA. This DNA is then used as a template to amplify a portion of the expected lentiviral insert. |
PRRSV is an infectious disease that causes lung damage and abortion in sows. Apoptosis of cells at the interface of the endometrium and placenta is an important factor leading to abortion. Previous studies have demonstrated that PRRSV can cause apoptosis of macrophages, but rarely causes obvious changes in porcine endometrial epithelial cells (PEC). Recently, abortion caused by PRRSV has been attributed to apoptosis of fetal placental and endometrial epithelial cells (Sn+ and CD163+). In this study, highly purified primary PECs were harvested by differential digestion and their characteristics were confirmed by CK18, ERɑ, and PR staining. The cells were then immortalized by transfection with a lentiviral vector expressing the SV40 large T antigen. PEC lines were obtained after puromycin selection. The proliferation of the cell lines was evaluated by cell growth curve and cell cycle analysis. The cell lines showed faster proliferation capacity than the primary cells. The biological characteristics of the cells were detected by Western blot, karyotype analysis, and staining, confirming that the cells retained endometrial characteristics. Finally, the sensitivity of PRRSV was tested, and the expression of Sn and CD163 indicated that both primary PECs and cell lines were potentially sensitive to PRRSV. The PRRSV infection test showed that the apoptosis rate of infected PEC cell lines increased significantly, indicating that the cell line was sensitive to PRRSV.
In this study, the SV40 large T antigen expression vector was transfected into primary PECs using packaged lentiviral particles to achieve cell immortalization. After puromycin selection, cells from a single cell clone showed significantly faster growth and higher adhesion efficiency compared with primary PECs. The PEC cell line was passaged for more than 50 generations while maintaining a homogenous population and unchanged properties. In addition, characteristic proteins found in primary PECs, such as CK18, ERɑ, and PR, could also be detected in the PEC cell line. In addition, the expression of ERɑ and PR was quantified by Western blotting. The results showed that the expression of these two proteins in PECs was not reduced compared with primary PECs. Therefore, the properties of PECs reconstructed by transfection are similar to those of primary PECs.
Figure 1. Characterization of the PEC cell line. (Zhang K, et al., 2019)
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