Does anyone know how to control the pH of the culture medium using the hypoxia incubator chamber? I'm using nitrogen to make the hypoxic condition and keeping the cells for 72h, but the culture medium is becoming alkaline (pinkish) and the cells are dying.
Hello Amanda,
I would recommend you to use HBSS in addition to media. That can be really useful. HBSS can easily handle issues related to CO2 fluctuations which ultimately result into PH change. Also the time 72 hrs is long. I was wondering if you can replace that with 48 hrs because media tends to deplete after 48 hrs if the cells are doing fine and metabolically active.
Hope it helps !
Hey, you might want to check your CO2 level. Usually media turns acidic through all the metabolites, especially when it is an incubation for 72h (I measured it once, and with my cells pH was changing within 48h.... 24h were fine). Pink usually means that you have a problem with your CO2. I dont know if you have a fairly old chamber (without the possibility of controlling CO2 - i still remember these old days). We had the problem that someone "fiddled" around with the software and changed basic levels of CO2 to a lot lower amount than 5%.
Well, I hope you can solve your problem. Good luck
Thank you, I'll try HBSS and 48h. I was using 72h because of the ideal time of action in a drug test. I didn't put CO2 Just N2. I'm using Billips-rothenberg modular incubator chamber.
To control and maintain the pH of the culture medium, the hypoxia incubator should be humidified and heated to body temperature. The humidity should be maintained by including an extra dish with sterile water. The increase in the alkalinity might be due to release of carbon dioxide in the atmosphere. You can control this by supplying carbon dioxide to the atmosphere, thereby preventing the carbon dioxide in the liquid from escaping, hence, the pH will not rise further.
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