CTS™ LV-MAX™ 生产培养基

No, CTS Viral Production Cells are HEK 293F suspension cells that have been adapted to CTS LV-MAX Production Medium. They do not express the SV40 large T antigen.

CTS Viral Production Cells can be frozen directly in cryogenic medium consisting of 90% CTS LV-MAX Production Medium plus 10% DMSO. Here is the protocol:

- Bank cells at passage 3 when cell density reaches ˜3.0-4.5 x 10E6 viable cells/mL at >95% viability.
- Centrifuge the cells at 100 x g (low speed) for 5 mins to pellet. Discard the spent medium.
- Prepare freezing medium: e.g., 10 mL of freezing medium is 9 mL of CTS LV-MAX Production Medium + 1 mL DMSO; mix well and sterilize by 0.2 µm filtration.
- Resuspend the cell pellet in the prepared freezing medium.
- Dilute the cells to a final density of 10 x10E6 viable cells/mL and aliquot 1 mL per cryovial.
- Freeze the cells at -80 degrees C for 1 day.
- Transfer frozen vials to liquid nitrogen for long-term storage.

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.