Cat.No. : AD00404Z
Titer: ≥1x10^10 IFU/mL / ≥1x10^11 IFU/mL / ≥1x10^11 VP/mL / ≥1x10^12 VP/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | AD00404Z |
| Description | Human Adenovirus Type5 (dE1/E3) expressing Vascular Endothelial Growth Factor with C-terminus 6xHN tag under a CMV promoter. C-terminus 6xHN tag, pre-made adenovirus, ready to ship and ready to use format. |
| Target Gene | VEGF |
| Product Type | Adenoviral particle |
| Insert | VEGF, C-fusion with 6xHN tag |
| Titer | Varies lot by lot, for example, ≥1x10^10 IFU/mL, ≥1x10^11 IFU/mL, ≥1x10^11 VP/mL etc. |
| Size | Varies lot by lot, for example, 250 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 adenovirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between adenovirus particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in adenovirus production, especially for applications in animal studies and gene therapy. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced adenovirus particles to ensure regulatory compliance. |
| Sterility | Creative Biogene ensures that adenovirus products are free of any bacterial, fungal and other microbial contamination. |
| Ad5 E1 Detection | All Creative Biogene adenoviruses are PCR tested to ensure that there are no detectable E1 sequences in the particles, which could be from revertants or external E1 contamination. |
| RCA Assays | Adenovirus products originating at Creative Biogene are guaranteed to have undetectable replication-competent adenovirus (RCA). This quality control measure is important because there is always the possibility of wild-type contamination due to revertants or environmental sources. |
| PFU Titering | All purified adenovirus preparations are tested for infectious titer. Creative Biogene's PFU test takes a few days longer but counts true plaques in HEK cells rather than estimating PFU titers via IHC staining or TCI50 of infected cells. |
| Gene Name | Vascular Endothelial Growth Factor |
| Gene Symbol | VEGF |
| Gene ID | 7422 |
| mRNA Refseq | NM_003376 |
Vascular endothelial growth factor (VEGF) stimulation and bone marrow mesenchymal stem cell (BMSC) transplantation have been used to treat acute cerebral infarction due to their key roles in behavioral recovery, neuroprotection, neurogenesis, and angiogenesis. However, the effect of BMSC transplantation may be limited due to the low graft survival rate after acute cerebral infarction, and the effect of VEGF delivery alone on recovery is also limited because the protein is rapidly cleared. Here, the researchers sought to explore whether VEGF can be transferred to BMSCs by adenovirus and whether transplantation of VEGF-transfected BMSCs into the rat brain can provide adequate neuroprotection after transient middle cerebral artery occlusion. Adenovirus carried VEGF into BMSCs (Ad-VEGF-BMSCs), and purified adenovirus was transferred into BMSCs (Ad-BMSCs). Rats exposed to 90-min middle cerebral artery occlusion (MCAO) were treated with Ad-VEGF-BMSC, Ad-BMSC, BMSC and Dulbecco's Modified Eagle's Medium (DMEM) after ischemia reperfusion for 24 h. After Ad-VEGF-BMSCs were transplanted into the local area of the ischemic rat brain tissue, it was found that the expression and secretion of VEGF and BDNF increased, the level of MAP2 increased, the microvascular density increased, the behavioral function improved, and the survival rate of BMSCs increased. The results showed that Ad-VEGF-BMSC transplantation can improve the ischemic neurological deficits after MCAO in rats, providing a potential valuable intervention for the treatment of cerebral ischemic diseases.
Fourteen days after transplantation, VEGF expression of transfected BMSCs in the transplanted area was detected by immunofluorescence staining and Western blot analysis (Figure 1A, B, and C). As shown in Figure 1A, some cells were positively stained for VEGF. Ad-VEGF-BMSCs have the potential to migrate from the BMSC transplantation site to the ischemic area. This result indicates that the transplanted BMSCs can survive in the ischemic area and that these BMSCs can functionally express VEGF protein in vivo. In addition, Ad-VEGF-BMSCs can continuously express VEGF protein, which may help promote neurogenesis and angiogenesis after MCAO. The VEGF expression in the Ad-VEGF-BMSC group was significantly higher than that in the other five groups, while the VEGF staining in the Sham group, MCAO group, and DMEM group was significantly lower than that in the BMSC group and the Ad-BMSC group. The VEGF protein level in the Ad-VEGF-BMSC group was significantly higher than that in the other five groups (Figure 1B, C), and the VEGF protein expression in the BMSC group and the Ad-BMSC group was significantly higher than that in the Sham group, the MCAO group, and the DMEM group. GFP-positive cell counts showed that the number of GFP-positive cells in the Ad-VEGF-BMSC group was significantly increased compared with that in the Ad-BMSC group (Figure 1D).
Figure 1. Immunofluorescence (IF) staining and Western blotting results. (Zong X, et al., 2017)
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The VEGF adenovirus achieved robust expression in our endothelial cell model. The virus was highly pure and the biological activity was consistent with the description. It is ideal for angiogenesis research!
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