ViroVision™ Infection Medium

ViroVision™ Infection Medium is a viral infection enhancer designed to facilitate viral penetration of the cortical actin barrier, thereby greatly enhancing productive viral infection. ViroVision™ Infection Medium can be used to facilitate the infection of a variety of host cells by different viruses and viral vectors. ViroVision™ Infection Medium can enhance viral infection rates by 5 to 20 fold. Virongy developed It is based on the scientific theory that the actin cytoskeleton is a natural barrier for viral entry and post-entry intracellular migration.

(Yoder et al., Cell, 2008, 134:782).

Applications

• Enhancing lenti- or retroviral transduction of target cells
• Enhancing infection rates of other enveloped viruses
• Facilitating recovery of infectious viruses from cell or tissue cultures
• Facilitating anti-viral drug screening efficiency

Table1:10 x Concentrated ViroVision™ Infection Medium

Protocol

Example 1 - ViroVision™ Infection Medium used to enhance lentiviral infection of suspension CEM-SS cells:

(See Table 2 for scale-up recommendations)

1. Count cells to be infected, and pellet cells by centrifugation at 300 x g for 5 minutes.

Note: Cell viability should be ≥ 80%.

2. Resuspend cells in complete media at a concentration of ~2 x 10⁶ cells mL^-1.
3. Use 100 μl of cells (~2 x 10⁵) per ViroVision™ Infection Medium.
4. Pre-treat cells by adding 10 μl of ViroVision™ Infection Medium so that ViroVision™ Infection Medium concentration is ~1X. Mix and incubate for 30–60 minutes.

For best results, when using ViroVision™ Infection Medium for the first time, start with a higher concentration (final ViroVision™ Infection Medium concentration may be 2X or 3X). The lowest effective dosage can be determined through a titration experiment (final ViroVision™ Infection Medium concentration may be as low as 1X).

5. Add virus to the cells and mix. Note volume of virus used.
6. Add ViroVision™ Infection Medium in an amount equal to 1/10 of the virus volume used, e.g., if 100 μl of virus is used, add 10 μl of ViroVision™ Infection Medium. Incubate at 37°C for 2-4 hours.
7. Add 1 ml fresh complete media to wash cells. Pellet cells as above and remove supernatant. Optional: Repeat once for a total of 2 washes.
8. After washing, resuspend cells in 1 ml complete medium.
9. Culture infected cells as usual to quantify viral replication.

Table 2: Scaleup Recommendations Using ViroVision™ Infection Medium

Example 2 - ViroVision™ Infection Medium enhances lentiviral transduction of adherent HDFn cells:

1. Count HDFn (Human Dermal Fibroblast, neonatal) cells to be infected and seed ~5 x 10⁵ cells per well into 6-well plates (2 ml per well).
2. Culture cells until cells stably adhere to the plates (4–12 hours).

Note: Cell viability should be ≥ 80%.

3. Before infection, wash cells with 2 ml medium, and leave 0.5 ml medium in each well.
4. Pre-treat cells by adding 50 μl of ViroVision™ Infection Medium so that Infectin concentration is ~1X. Mix and incubate for 30–60 minutes.

For best results, when using ViroVision™ Infection Medium for the first time, start with a higher concentration (final Infectin concentration may be 2X or 3X). The lowest effective dosage can be determined through a titration experiment (final ViroVision™ Infection Medium concentration may be as low as 1X).

5. Add virus to cells and mix. Note volume of virus used.
6. Add ViroVision™ Infection Medium in an amount equal to 1/10 of the virus volume used, e.g., if 100 μl of virus is used, add 10 μl of ViroVision™ Infection Medium. Incubate at 37°C for 2–4 hours.
7. Add 2 ml fresh media to wash cells.
8. After washing, add 2 ml fresh complete medium.
9. Culture infected cells as usual.

Viral Enhancement Results:

ViroVision™ Infection Medium enhances lentiviral transduction of Hep G2 cells:

• • Hep G2 cells were transduced with lenti-GFP vector in the presence or absence of ViroVision™ Infection Medium.