Transparent polymeric cell culture chip with integrated temperature control and uniform media perfusion.
Modern microfabrication and microfluidic technologies offer new opportunities in the design and fabrication of miniaturized cell culture systems for online monitoring of living cells. We used laser micromachining and thermal bonding to fabricate an optically transparent, low-cost polymeric chip for long-term online cell culture observation under controlled conditions. The chip incorporated a microfluidic flow equalization system, assuring uniform perfusion of the cell culture media throughout the cell culture chamber. The integrated indium-tin-oxide heater and miniature temperature probe linked to an electronic feedback system created steady and spatially uniform thermal conditions with minimal interference to the optical transparency of the chip. The fluidic and thermal performance of the chip was verified by finite element modeling and by operation tests under fluctuating ambient temperature conditions. HeLa cells were cultured for up to 2 weeks within the cell culture chip and monitored using a time-lapse video recording microscopy setup. Cell attachment and spreading was observed during the first 10-20 h (lag phase). After approximately 20 h, cell growth gained exponential character with an estimated doubling time of about 32 h, which is identical to the observed doubling time of cells grown in standard cell culture flasks in a CO2 incubator.
Petronis
Sarunas
S
Technical University of Denmark, Lyngby.
Stangegaard
Michael
M
Christensen
Claus Bo Vöge
CB
Dufva
Martin
M
D018929:Cell Culture Techniques / Q000295:instrumentation*; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D017076:Computer-Aided Design; D003470:Culture Media / Q000378:metabolism; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000379:methods*; D006367:HeLa Cells; D006361:Heating / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D018715:Microscopy, Video / Q000379:methods*; D019904:Polymethyl Methacrylate; D018511:Systems Integration; D013696:Temperature
D003470:Culture Media; D019904:Polymethyl Methacrylate