Sf9 insect cell line. Sf9 cells come from ovarian tissue of the fall armyworm. The advantages of using these cells are that the incubation temperature is close to room temperature (26-28°C) and CO2 is not required. This worked out nicely because our CO2 incubator was still on order.
We used an Edvotek kit which came with an OptiCell chamber with the live insect cells inside. Each student got to inoculate a flask in the biological safety cabinet. It was a great way to practice aseptic technique. Since the semester ended, I have been passaging their cultures and trying different media. The formulation that seems to work best is Grace’s Insect Media, supplemented (TNM-FH, Lonza) with 10% heat-inactivated fetal bovine serum (FBS). I have also tried Thermo Scientific HyClone SFX Insect Media (Thermo Scientific HyClone).
Here are some pictures of the results. By the way, the inverted scope we used was found in pieces in a box in the basement of our building. It appears to be a Nikon circa 1986. Some students and I put it back together and it works fine.
The next steps will be scaling up to spinner flasks for our two sizes of bioreactors and then transfection with baculovirus!