I'm wondering if anyone has insight into the effect of minimal changes in CO2 incubator levels on primary neuronal cell cultures? Cells are plated at a density of 4.66104 cells per cm2 in 384-well poly-D Lysine coated plates in Neurobasal Media, supplemented with B27,Glutamax and antibiotics (penicillin, 50 units/ml and streptomycin 50 mg/ml). Cultures are typically maintained at 37C in 5% CO 2. When reading the CO2 levels with fyrite, we get fluctuation with readings of 5.5% to 6% CO2. Could this cause cell stress?
Does your media contain phenol red? Do you observe a color change? For a small fluctuation like that, I would say the biggest concern is decreasing pH of your culture media. However, I think your NB is phosphate and HEPEs buffered, so small fluctuations shouldn't be a big deal. It's common to see fluctuations from 5% down to 4.0% just by opening the incubator door. Additionally, neurons are relatively insensitive to extracellular acidification. However, if these fluctuations are more than transitory in duration, you may be introducing variability into whatever experiments you are trying.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning