CTS™ Opti-MEM™ I Medium

We recommend using V-bottom 96-well plates to harvest the supernatant containing lentiviral particles as they allow for easier detection of the cell pellets after centrifugation of the cultures.

We recommend maintaining shaking cultures in a separate incubator from all other incubators to avoid vibration effects. Alternatively, we recommend using a small, quiet shaker such as the Thermo Scientific CO₂ Resistant Shaker (Cat. No. 88881102) for convenience and best performance.

We generally recommend culturing the CTS Viral Production Cells under 8% CO₂ conditions. We found that cells cultured in 5% CO₂ grew slower and were more prone to more cell clumping than when grown in 8% CO₂.

The CTS LV-MAX lentivirus production system is a fully-optimized complete system in which the cells, media, and transfection reagent work together to generate high-titer lentivirus. The CTS LV-MAX Production Medium is specifically designed to work with the system, to support high density suspension cell growth of CTS Viral Production Cells.

Although 3-4 passages before transfection are sufficient for most cell lines recovering directly from liquid N2 storage, we found that CTS Virus Production Cells passaged to P5 showed the highest performance in lentivirus particle production and reproducibility between batches. Therefore, we recommend passaging the cells to P4 at the end of a given week for lentivirus production the following week.