On-chip CO ₂ incubation for pocket-sized microfluidic cell culture

We have developed an on-chip CO ₂ incubation system based on mass/heat transfer from aqueous solutions of bicarbonate source to cell culture media through a permeable poly(dimethylsiloxane) (PDMS) wall. Heating a carbonate-buffered bicarbonate solution successfully regulated CO ₂ generation without any feedback control. Because a microfluidic cell culture chip with the incubation system does not require an external chamber or gas supply, the entire microfluidic cell culture setup becomes pocket sized. Using 5 ml of 0.8 M sodium bicarbonate with 65 mM sodium carbonate as the water jacket, the chip maintained the temperature, osmolality, and pH of 750 μl cell culture medium within physiological levels when the chip was placed on a 37°C surface. The osmolality shift and pCO ₂ of the media reservoir stabilized within <5 mmol/kg and 5.0 ± 1.0% over at least 9 days. The incubation capabilities were demonstrated through microfluidic culture of COS-7 epithelial cells under an inverted microscope for 17 days.